Methods and materials for fabricating microfluidic devices

ABSTRACT

Materials and Methods are provided for fabricating microfluidic devices. The materials include low surface energy fluoropolymer compositions having multiple cure functional groups. The materials can include multiple photocurable and/or thermal-curable functional groups such that laminate devices can be fabricated. The materials also substantially do not swell in the presence of hydrocarbon solvents.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is based on and claims priority to U.S. ProvisionalPatent Application Ser. No. 60/706,786, filed Aug. 9, 2005; U.S.Provisional Patent Application Ser. No. 60/732,727, filed Nov. 2, 2005;and U.S. Provisional Patent Application Ser. No. 60/799,317 filed May10, 2006; each of which is incorporated herein by reference in itsentirety.

This application is also a continuation-in-part of PCT InternationalPatent Application Serial No. PCT/US05/04421, filed Feb. 14, 2005, whichis based on and claims priority to U.S. Provisional Patent ApplicationSer. No. 60/544,905, filed Feb. 13, 2004, each of which is incorporatedherein by reference in its entirety.

GOVERNMENT INTEREST

This invention was made with U.S. Government support from Office ofNaval Research No. N000140210185 and STC program of the National ScienceFoundation under Agreement No. CHE-9876674. The U.S. Government hascertain rights in the invention.

TECHNICAL FIELD

Generally, the present invention relates to materials and methods forfabricating polymeric devices. More particularly the polymeric devicescan be multi-layered devices such as microfluidic devices.

ABBREVIATIONS

-   -   AC=alternating current    -   Ar=Argon    -   ° C.=degrees Celsius    -   cm=centimeter    -   8-CNVE=perfluoro(8-cyano-5-methyl-3,6-dioxa-1-octene)    -   CSM=cure site monomer    -   CTFE=chlorotrifluoroethylene    -   g=grams    -   h=hours    -   1-HPFP=1,2,3,3,3-pentafluoropropene    -   2-HPFP=1,1,3,3,3-pentafluoropropene    -   HFP=hexafluoropropylene    -   HMDS=hexamethyldisilazane    -   IL=imprint lithography    -   IPDI=isophorone diisocyanate    -   MCP=microcontact printing    -   Me=methyl    -   MEA=membrane electrode assembly    -   MEMS=micro-electro-mechanical system    -   MeOH=methanol    -   MIMIC=micro-molding in capillaries    -   mL=milliliters    -   mm=millimeters    -   mmol=millimoles    -   M_(n)=number-average molar mass    -   m.p.=melting point    -   mW=milliwatts    -   NCM=nano-contact molding    -   NIL=nanoimprint lithography    -   nm=nanometers    -   Pd=palladium    -   PAVE perfluoro(alkyl vinyl)ether    -   PDMS=poly(dimethylsiloxane)    -   PEM=proton exchange membrane    -   PFPE=perfluoropolyether    -   PMVE perfluoro(methyl vinyl)ether    -   PPVE perfluoro(propyl vinyl)ether    -   PSEPVE=perfluoro-2-(2-fluorosulfonylethoxy)propyl vinyl ether    -   PTFE=polytetrafluoroethylene    -   SAMIM=solvent-assisted micro-molding    -   SEM=scanning electron microscopy    -   Si=silicon    -   TFE=tetrafluoroethylene    -   μm=micrometers    -   UV=ultraviolet    -   W=watts    -   ZDOL=poly(tetrafluoroethylene oxide-co-difluoromethylene        oxide)α,ω diol

BACKGROUND

Microfluidic devices developed in the early 1990s were fabricated fromhard materials, such as silicon and glass, using photolithography andetching techniques. See Ouellette, J., The Industrial Physicist 2003,Aug./Sep., 14-17; Scherer, A., et al., Science 2000, 290, 1536-1539.Photolithography and etching techniques, however, are costly and laborintensive, require clean-room conditions, and pose several disadvantagesfrom a materials standpoint. For these reasons, soft materials haveemerged as alternative materials for microfluidic device fabrication.The use of soft materials has made possible the manufacture andactuation of devices containing valves, pumps, and mixers. See, e.g.,Ouellette, J., The Industrial Physicist 2003, Aug./Sep., 14-17; Scherer,A., et al., Science 2000, 290, 1536-1539; Unger, M. A., et al., Science2000, 288, 113-116; McDonald, J. C., et al., Acc. Chem. Res. 2002, 35,491-499; and Thorsen, T., et al., Science 2002, 298, 580-584. Forexample, one such microfluidic device allows for control over flowdirection without the use of mechanical valves. See Zhao, B., et al.,Science 2001, 291, 1023-1026.

The increasing complexity of microfluidic devices has created a demandto use such devices in a rapidly growing number of applications. To thisend, the use of soft materials has allowed microfluidics to develop intoa useful technology that has found application in genome mapping, rapidseparations, sensors, nanoscale reactions, ink-jet printing, drugdelivery, Lab-on-a-Chip, in vitro diagnostics, injection nozzles,biological studies, and drug screening. See, e.g., Ouellette, J., TheIndustrial Physicist 2003, Aug./Sep., 14-17; Scherer, A., et al.,Science 2000, 290, 1536-1539; Unger, M. A., et al., Science 2000, 288,113-116; McDonald, J. C., et al., Acc. Chem. Res. 2002, 35, 491-499;Thorsen, T., et al., Science 2002, 298, 580-584; and Liu, J., et al.,Anal. Chem. 2003, 75, 4718-4723.

Poly(dimethylsiloxane) (PDMS) is the soft material of choice for manymicrofluidic device applications. See Scherer, A., et al., Science 2000,290, 1536-1539; Unger, M. A., et al., Science 2000, 288, 113-116;McDonald, J. C., et al., Acc. Chem. Res. 2002, 35, 491-499; Thorsen, T.,et al., Science 2002, 298, 580-584; and Liu, J., et al., Anal. Chem.2003, 75, 4718-4723. A PDMS material offers numerous attractiveproperties in microfluidic applications. Upon cross-linking, PDMSbecomes an elastomeric material with a low Young's modulus, e.g.,approximately 750 kPa. See Unger, M. A., et al., Science 2000, 288,113-116. This property allows PDMS to conform to surfaces and to formreversible seals. Further, PDMS has a low surface energy, e.g.,approximately 20 erg/cm², which can facilitate its release from moldsafter patterning. See Scherer, A., et al., Science 2000, 290, 1536-1539;McDonald, J. C., et al., Acc. Chem. Res. 2002, 35, 491-499.

Another important feature of PDMS is its outstanding gas permeability.This property allows gas bubbles within the channels of a microfluidicdevice to permeate out of the device. This property also is useful insustaining cells and microorganisms inside the features of themicrofluidic device. The nontoxic nature of silicones, such as PDMS,also is beneficial in this respect and allows for opportunities in therealm of medical implants. McDonald, J. C., et al., Acc. Chem. Res.2002, 35, 491-499.

Many current PDMS microfluidic devices are based on SYLGARD® 184 (DowCorning, Midland, Mich., United States of America). SYLGARD® 184 iscured thermally through a platinum-catalyzed hydrosilation reaction.Complete curing of SYLGARD® 184 can take as long as five hours. Thesynthesis of a photocurable PDMS material, however, with mechanicalproperties similar to that of SYLGARD® 184 for use in soft lithographyrecently has been reported. See Choi, K. M., et al., J. Am. Chem. Soc.2003, 125, 4060-4061.

Despite the aforementioned advantages, PDMS suffers from multipledrawbacks in microfluidic applications. First, PDMS swells in mostorganic solvents. Thus, PDMS-based microfluidic devices have a limitedcompatibility with various organic solvents. See Lee, J. N. et al.,Anal. Chem. 2003, 75, 6544-6554. Among those organic solvents that swellPDMS are hexanes, ethyl ether, toluene, dichloromethane, acetone, andacetonitrile. See Lee, J. N., et al., Anal. Chem. 2003, 75, 6544-6554.The swelling of a PDMS microfluidic device by organic solvents candisrupt its micron-scale features, e.g., a channel or plurality ofchannels, and can restrict or completely shut off the flow of organicsolvents through the channels. Thus, microfluidic applications with aPDMS-based device are limited to the use of fluids, such as water, thatdo not swell PDMS. As a result, those applications that require the useof organic solvents likely will need to use microfluidic systemsfabricated from hard materials, such as glass and silicon. See Lee, J.N., et al., Anal. Chem. 2003, 75, 6544-6554. This approach, however, islimited by the disadvantages of fabricating microfluidic devices fromhard materials.

Second, PDMS-based devices and materials are notorious for not beingadequately inert to be used even in aqueous-based chemistries. Forexample, PDMS is susceptible to reaction with weak and strong acids andbases. PDMS-based devices also are notorious for containingextractables, such as extractable oligomers and cyclic siloxanes,especially after exposure to acids and bases. Because PDMS is easilyswollen by organics, hydrophobic materials, even those hydrophobicmaterials that are slightly soluble in water, can partition during useinto PDMS-based materials used to construct PDMS-based microfluidicdevices.

Thus, an elastomeric material that exhibits the attractive mechanicalproperties of PDMS combined with a resistance to swelling in commonorganic solvents would extend the use of microfluidic devices to avariety of new chemical applications that are inaccessible by currentPDMS-based devices. Accordingly, the approach demonstrated by thepresently disclosed subject matter uses an elastomeric material, moreparticularly a functional perfluoropolyether (PFPE) material, which isresistant to swelling in common organic solvents to fabricate amicrofluidic device.

Functional PFPE materials are liquids at room temperature, exhibit lowsurface energy, low modulus, high gas permeability, and low toxicitywith the added feature of being extremely chemically resistant. SeeScheirs, J., Modern Fluoropolymers; John Wiley & Sons, Ltd.: New York,1997; pp 435-485. Further, PFPE materials exhibit hydrophobic andlyophobic properties. For this reason, PFPE materials are often used aslubricants on high-performance machinery operating in harsh conditions.The synthesis and solubility of PFPE materials in supercritical carbondioxide has been reported. See Bunyard, W., et al., Macromolecules 1999,32, 8224-8226. Beyond PFPEs, fluoroelastomers also can includefluoroolefin-based materials, including, but not limited to, copolymersof tetrafluoroethylene, hexafluoropropylene, vinylidene fluoride andalkyl vinyl ethers, often with additional cure site monomers added forcrosslinking.

A PFPE microfluidic device has been previously reported by Rolland, J.et al. JACS 2004, 126, 2322-2323. The device was fabricated from afunctionalized PFPE material (e.g., a PFPE dimethacrylate (MW=4,000g/mol)) having a viscosity of the functionalized material ofapproximately 800 cSt. This material was end-functionalized with a freeradically polymerizable methacrylate group and UV photocured freeradically with a photoinitiator. In Rolland, J. et al., supra,multilayer PFPE devices were generated using a specific partial UVcuring technique, however, the adhesion was weak and generally notstrong enough for a wide range of microfluidic applications. Further,the adhesion technique described by Rolland, J. et al. did not providefor adhesion to other substrates such as glass.

The presently disclosed subject matter describes the use offluoroelastomers, especially a functional perfluoropolyether as amaterial for fabricating a solvent-resistant micro- and nano-scalestructures, such as a microfluidic device. The use of fluoroelastomersand functional perfluoropolyethers in particular as materials forfabricating a microfluidic device addresses the problems associated withswelling in organic solvents exhibited by microfluidic devices made fromother polymeric materials, such as PDMS. Accordingly, PFPE-basedmicrofluidic devices can be used in conjunction with chemical reactionsthat are not amenable to other polymeric microfluidic devices.

Further, adequate adhesion between the layers of a multilayermicrofluidic device can impact performance in most if not allapplications. For example, adhesion can impact performance inmicrofluidic devices, such as those described by Unger et al., Science,288, 113-6 (2000), which contain multiple layers allowing for theformation of pneumatic valves. Thus, there is a need in the art forimproved methods for adhering the layers of a microfluidic devicetogether or to other surfaces.

Furthermore, many devices, such as surgical instruments, medicaldevices, prosthetic implants, contact lenses, and the like, are formedfrom polymeric materials. Polymeric materials commonly used in themedical device industry include, but are not limited to polyurethanes,polyolefins (e.g., polyethylene and polypropylene), poly(meth)acrylates,polyesters (e.g., polyethyleneterephthalate), polyamides, polyvinylresins, silicone resins (e.g., silicone rubbers and polysiloxanes),polycarbonates, polyfluorocarbon resins, synthetic resins, polystyrene,various bioerodible materials, and the like. Although these and othermaterials commonly used as implant materials have proven to be usefulthere are many drawbacks with the materials. One drawback is that withany implant there is always the chance of bio-fouling on the surface ofthe implant. Bio-fouling can occur due to the tissue/implant interfacegap and the surface characteristics of the implant material.Accordingly, a need exists for improving the polymeric materials and/orfunctionalizing the materials or the surface of the medical devicematerials to generate a better tissue/device interface and reducebio-fouling.

SUMMARY

According to an embodiment of the present invention, a polymercomposition includes a first component including a fluoropolymer havinga first curable functional group and a second component including afluoropolymer having a second curable functional group. In someembodiments, the composition further includes a third componentincluding a fluoropolymer having a third curable functional group.According to some embodiments, at least one of the fluoropolymer havinga first curable functional group or the fluoropolymer having a secondcurable functional group comprises a perfluoropolyether. In alternativeembodiments, at least one of the fluoropolymer having a first curablefunctional group, the fluoropolymer having a second curable functionalgroup, or the fluoropolymer having a third curable functional groupcomprises a low surface energy polymeric material.

In some embodiments, the first curable functional group includes a firstphotocurable functional group. In some embodiments, the firstphotocurable functional group includes a photocurable diurethanemethacrylate. In some embodiments, the second curable functional groupincludes a second photocurable functional group, which can include aphotocurable diepoxy.

According to some embodiments, the second curable functional groupincludes a first thermal-curable functional group. In some embodiments,the first thermal-curable functional group includes a diisocyanate, adiepoxy, or a diamine. In some embodiments, the third curable functionalgroup includes a second thermal-curable functional group that includes atriol, or a tetrol. The composition can include the combination whereinthe first curable functional group includes a photocurable diurethanemethacrylate, the second curable functional group includes adiisocyanate, and the third curable functional group includes a triol.According to other embodiments, the composition can include thecombination wherein the first curable functional group includes aphotocurable diurethane methacrylate, the second curable functionalgroup includes a diisocyanate, and the third curable functional groupincludes a tetrol. In other embodiments, the composition can include thecombination wherein the first curable functional group includes aphotocurable diurethane methacrylate, the second curable functionalgroup includes a diepoxy, and the third curable functional groupincludes a diamine. In still further embodiments, the composition caninclude the combination wherein the first curable functional groupincludes a photocurable diurethane methacrylate and the second curablefunctional group includes a photocurable diepoxy. In furtherembodiments, composition can include the combination wherein the firstcurable functional group includes a photocurable diurethane methacrylateand the second curable functional group includes a diisocyanate.According to some embodiments, the material of the present invention caninclude a fluoropolymer fabricated from a fluoroalkyliodide precursor.

According to some embodiments, the first photocurable functional groupreacts at a first wavelength and the second photocurable functionalgroup react at a second wavelength. In other embodiments, the firstthermal-curable functional group reacts at a first time period and thesecond thermal-curable functional group reacts at a second time period.According to other embodiments, the first thermal-curable functionalgroup reacts at a first temperature and the second thermal-curablefunctional group reacts at a second temperature. According to someembodiments, the second curable functional group remains viable afterthe first curable functional group is activated such that the secondcurable functional group can bind with a group consisting of anotherpolymer, a hydroxyl group, another functional group, and combinationsthereof.

In some embodiments, the perfluoropolyether is encapped withcycloaliphatic epoxides moieties. In some embodiments, theperfluoropolyether has a molecular weight of about 16000 and a modulusof about 800 kPa. In other embodiments, the perfluoropolyether has amolecular weight of less than about 16000. In further embodiments, theperfluoropolyether has a modulus of greater than about 500 kPa. In otherembodiments, the perfluoropolyether has a molecular weight of about16000 and a percent elongation at break of about 200 percent. Accordingto some embodiments, the perfluoropolyether has a percent elongation atbreak of less than about 200 percent. In other embodiments, one of thefirst component or the second component includes a fluoropolymer havingan elongation at break of about 300 percent. In yet other embodiments,one of the first component or the second component includes afluoropolymer having an elongation at break of about 200 percent. Inother embodiments, one of the first component or the second componentincludes a fluoropolymer having an elongation at break of between about100 percent and about 300 percent. According to some embodiments, one ofthe first component or the second component includes aperfluoropolyether having an elongation at break of between about 200percent and about 300 percent.

According to some embodiments, a polymer composition includes afluoropolymer having at least two functional groups, wherein after afirst functional group is activated to polymerize the fluoropolymer, asecond functional group remains activatable to adhere the fluoropolymerto another chemical group. In some embodiments, the another chemicalgroup is selected from the group of another polymer, a hydroxyl group,another functional group, and combinations thereof.

According to some embodiments of the present invention, a microfluidicdevice includes a first fluoropolymer component, wherein the firstfluoropolymer component includes a perfluoropolyether. In someembodiments, the first fluoropolymer component includes a first curablefunctional group and can also include a second fluoropolymer componenthaving a second curable functional group. In some embodiments, the firstcurable functional group includes a first photocurable functional groupand the second curable functional group includes a first thermal-curablefunctional group. According to other embodiments, the second curablefunctional group includes a second photocurable functional group.

In some embodiments, the first photocurable functional group includes aphotocurable diurethane methacrylate, or a photocurable diepoxy. In someembodiments, the first thermal-curable functional group includes adiisocyanate, a diepoxy, or a diamine. In further embodiments, themicrofluidic device further includes a third fluoropolymer component,wherein the third fluoropolymer component includes a third curablefunctional group. In some embodiments, the third curable functionalgroup includes a second thermal-curable functional group such as a triolor a tetrol.

According to some embodiments, the microfluidic device includes acomposition wherein the first curable functional group includes aphotocurable diurethane methacrylate, the second curable functionalgroup includes a diisocyanate, and the third curable functional groupincludes a triol. According to some embodiments, the microfluidic deviceincludes a composition wherein the first curable functional groupincludes a photocurable diurethane methacrylate, the second curablefunctional group includes a diisocyanate, and the third curablefunctional group includes a tetrol. According to some embodiments, themicrofluidic device includes a composition wherein the first curablefunctional group includes a photocurable diurethane methacrylate, thesecond curable functional group includes a diepoxy, and the thirdcurable functional group includes a diamine. According to someembodiments, the microfluidic device includes a composition wherein thefirst curable functional group includes a photocurable diurethanemethacrylate and the second curable functional group includes aphotocurable diepoxy. According to some embodiments, the microfluidicdevice includes a composition wherein the first curable functional groupincludes a photocurable diurethane methacrylate and the second curablefunctional group includes a diisocyanate. According to some embodiments,the microfluidic device includes a composition wherein the first curablefunctional group comprises a first photocurable functional group, andwherein the first photocurable functional group and the secondphotocurable functional group react to different wavelengths. Accordingto some embodiments, the microfluidic device includes a compositionwherein at least one of the first fluoropolymer component, the secondfluoropolymer component, and the third fluoropolymer component comprisesa perfluoropolyether. According to some embodiments, the microfluidicdevice includes a composition wherein the first fluoropolymer component,the second fluoropolymer component, and the third fluoropolymercomponent comprises a perfluoropolyether.

In other embodiments, the microfluidic device includes a siloxanecomponent coupled with a first fluoropolymer component. In someembodiments, the microfluidic device has a first fluoropolymer componentcoupled with a siloxane component forming an adhesion that can withstandup to about 120 pounds per square inch (psi). In some embodiments, thefirst fluoropolymer component of the microfluidic device has anelongation at break of about 300 percent. In other embodiments, thefirst fluoropolymer component has an elongation at break of about 200percent. In further embodiments, the first fluoropolymer component hasan elongation at break of between about 100 percent and about 300percent. In yet further embodiments, the first fluoropolymer componenthas an elongation at break of between about 200 percent and about 300percent. In other embodiments, the fluoropolymer includes a low surfaceenergy fluoropolymer. In some embodiments, the low surface energyfluoropolymer includes a surface energy of less than about 20 dynes/cm.In other embodiments, the low surface energy fluoropolymer includes asurface energy of less than about 18 dynes/cm, less than about 15dynes/cm, or less than about 12 dynes/cm.

According to some embodiments, the microfluidic device includes acomponent selected from the group of: diurethane methacrylate;chain-extended diurethane methacrylate; diisocyanate; chain extendeddiisocyanate; blocked diisocyanate; PFPE three-armed triol; PFPEdistyrene; diepoxy; diamine; thermally cured PU-tetrol; thermally curedPU-triol; thermally cured epoxy; photocured epoxy, and combinationsthereof.

In some embodiments, the microfluidic device includes a firstfluoropolymer component that is substantially free of trace metals. Inother embodiments, the first fluoropolymer component is substantiallysolvent resistant. In some embodiments, the first fluoropolymercomponent swells less than about 10% by weight when in communicationwith an organic solvent. In other embodiments, the fluoropolymercomponent swells less than about 5% by weight when in communication withan organic solvent.

In some embodiments, the fluoropolymer component has a carbon tofluorine ratio of about 1:2, while in other embodiments, thefluoropolymer component has a carbon to fluorine ratio of about 1:1.According to some embodiments, the fluoropolymer component has a carbonto fluorine ratio of about 2:1. In further embodiments, thefluoropolymer component has a carbon to fluorine ratio of between about1:1 and about 2:1. According to yet further embodiments, thefluoropolymer component has a carbon to fluorine ratio of between about1:1 and about 1:2.

According to some embodiments, the microfluidic device further includeslamination between a first part and a second part wherein the laminateincludes a chemical bond between the first part and the second part andwherein the first part does not delaminate from the second part under apressure of about 120 pounds per square inch. In some embodiments, themicrofluidic device includes a first part defining a channel therein anda valve positioned within the channel, wherein the valve can be actuatedat a pressure between about 40 psi and about 60 psi without damaging themicrofluidic device. In some embodiments, the microfluidic deviceincludes a first fluoropolymer component including a coating on achannel of the microfluidic device.

In some embodiments, the microfluidic device includes a firstfluoropolymer component having a molecular weight of about 16000 and amodulus of about 800 kPa. In other embodiments, the first fluoropolymercomponent has a molecular weight of less than about 16000. In someembodiments, the first fluoropolymer component has a modulus of greaterthan about 500 kPa. In further embodiments, the first fluoropolymercomponent has a molecular weight of about 16000 and a percent elongationat break of about 200 percent. In some embodiments, the firstfluoropolymer component has a percent elongation at break of less thanabout 200 percent.

According to some embodiments, a microfluidic device includes aperfluoropolyether epoxy-containing a PAG; wherein the combination isblended with about 1 to about 5 mole % of a free radical photoinitiatorselected from the group consisting of 2,2-dimethoxyacetophenone,1-hydroxy cyclohexyl phenyl ketone, or diethoxyacetophenone. In otherembodiments, the microfluidic device includes a perfluoropolyetherepoxy-containing a PAG is photocurable at two or more wavelengths. Insome embodiments, a microfluidic device includes a first part configuredfrom a perfluoropolyether and a second part configured from a silicone.According to an embodiment of the present invention a micro device isprepared by a process including treating the device with a solutioncomprising 0.5% Fluorine gas in Nitrogen such that the Fluorine reactsfree radically with hydrogen atoms in the device and thereby passivatesa surface of the device.

According to some embodiments, a microfluidic device is prepared by theprocess including adding a fluorinated fluid to a polymer precursor ofthe microfluidic device, wherein the polymer precursor includes aphotocurable or a thermalcurable precursor, curing the polymer bytreating the polymer with photo radiation or thermal energy,respectively, and removing the fluorinated fluid from the cured polymer.In some embodiments the removing is an evaporation process, or adissolving process. According to alternative embodiments, aconcentration of the fluorinated fluid is less than about 15%, less thanabout 10%, or less than about 5%.

In some embodiments, a micro device of the present invention includes asurface passivated with an end-capped precursor. In some embodiments,the end-capped precursor includes a styrene end-capped liquid precursor.In alternative embodiments, the surface includes a surface of a valve, achannel, a reservoir, a membrane, or a wall.

According to alternative embodiments of the present invention, componentpart of a microfluidic device include valves, membranes, channels,reservoirs, wells, lids, and the like. In some embodiments, thecomponent parts are fabricated from a first polymer component, whereinthe first polymer component includes a fluoropolymer having a firstcurable functional group and a second polymer component, wherein thesecond polymer component includes a fluoropolymer having a secondcurable functional group. In some embodiments, the component partincludes a third polymer component, wherein the third polymer componentincludes a fluoropolymer having a third curable functional group.According to some embodiments, the component part includes a compositionwherein at least one of the fluoropolymer having a first curablefunctional group, the fluoropolymer having a second curable functionalgroup, or the fluoropolymer having a third curable functional groupcomprises a perfluoropolyether. In other embodiments, the component partincludes a composition wherein at least one of the fluoropolymer havinga first curable functional group, the fluoropolymer having a secondcurable functional group, or the fluoropolymer having a third curablefunctional group comprises a low surface energy polymeric material. Insome embodiments, the fluoropolymer has a first curable functional groupincludes a photocurable functional group. In some embodiments, thecomponent part includes a fluoropolymer having a second curablefunctional group includes a thermal-curable functional group. In otherembodiments, the component part includes a photocurable functional groupselected from the group of photocurable diurethane methacrylate,photocurable diepoxy, and combinations thereof. In some embodiments, thethermal-curable functional group can be triol, diisocyanate, tetrol,diepoxy, diamine, or combinations thereof. In other embodiments, thefirst curable functional group and the second curable functional groupis selected from the group consisting of diurethane methacrylate;chain-extended diurethane methacrylate; diisocyanate; chain extendeddiisocyanate; blocked diisocyanate; PFPE three-armed triol; PFPEdistyrene; diepoxy; diamine; thermally cured PU-tetrol; thermally curedPU-triol; thermally cured epoxy; and photo-cured epoxy, and combinationsthereof. In some embodiments, the component part includes a membrane orvalve having at least two layer, wherein the layers include aperfluoropolyether laminate having a distyrene material laminated to aperfluoropolyether diurethane methacrylate material.

In some embodiments the component part includes a fluoropolymer havingpores defined therein. In alternative embodiments, the pores are lessthan about 15 percent, less than about 10 percent, or less than about 5percent.

According to some embodiments of the present invention, a method ofmaking a micro device includes fabricating a microfluidic device from afluoropolymer. In some embodiments, the fluoropolymer includesperfluoropolyether. In other embodiments, the fluoropolymer includes afirst fluoropolymer component having a first curable functional groupand a second fluoropolymer component having a second curable functionalgroup. According to some embodiments, the first curable functional groupincludes a photocurable functional group and the second curablefunctional group includes a thermal-curable functional group.

According to some embodiments, a method of making a microfluidic deviceincludes removing trace metals from a polymer and using the polymer tofabricate a microfluidic device, wherein the microfluidic device issubstantially free from trace metals. In some embodiments, the polymerincludes a fluoropolymer and in some embodiments, the fluoropolymerincludes perfluoropolyether.

According to some embodiments of the present invention, a method forconducting reactions includes providing a microfluidic device having apolymer substantially free from trace metals and conducting reactionsinvolving F— in the microfluidic device such that the F— issubstantially not quenched by trace metals in the polymer of themicrofluidic device.

In some embodiments of the present invention, a method for fabricating amicrofluidic device includes casting a fluoropolymer onto a mastertemplate, wherein the master template includes a dimensional pattern,curing the fluoropolymer such that the fluoropolymer retains asubstantial mirror image of the dimensional pattern, casting apoly(dimethylsiloxane) onto the cured fluoropolymer, curing thepoly(dimethylsiloxane) such that the poly(dimethylsiloxane) retains asubstantial mirror image of the pattern of the cured fluoropolymer, andusing the cured poly(dimethylsiloxane) as a mold for molding furtherfluoropolymer components.

According to other embodiments of the present invention, a method forfabrication of a microfluidic device includes spin coating a thin layerof uncured polymer onto a first portion of a microfluidic device,positioning the first portion on a second portion of the microfluidicdevice such that the spin coated thin layer is in communication with thesecond portion of the microfluidic device, and curing the combinationsuch that the spin coated thin layer is cured.

According to some embodiments, a method of increasing chemicalcompatibility of a polymeric device includes treating a polymeric devicecomprising a latent methacrylate, acrylate, and/or styrene group with asolution comprising a styrene end-capped precursor solution, evaporatingor dissolving the solution such that a film of the styrene end-cappedprecursor remains on a surface of the polymeric device, and curing thestyrene end-capped precursor film such that the film adheres to thesurface of the polymeric device through reaction with latentmethacrylate, acrylate, and/or styrene groups of the polymeric device.

In some embodiments of the present invention, a method of using a microdevice includes fabricating a microfluidic device from a fluoropolymer,wherein the microfluidic device includes a channel and flowing a liquidat least partially through the channel. In some embodiments, the liquidincludes an organic solvent, such as a hydrocarbon solvent. Inalternative embodiments, the hydrocarbon solvent is selected from thegroup of tetrahydrofuran, toluene, dichloromethane, hexanes, chloroform,isopropyl alcohol, cyclohexane, methyl ethyl ketone, acetone, andcombinations thereof. According to other methods of using a microfluidicdevice, a method is provided where the microfluidic device is fabricatedfrom a fluoropolymer that is substantially free of trace metals andwherein the liquid includes F—. In alternative embodiments, a method ofusing a microfluidic device includes providing a microfluidic devicehaving a channel, wherein the channel is partially coated with afluoropolymer and introducing a substance into the channel of themicrofluidic device, wherein the substance includes an organic solventsuch as a hydrocarbon solvent.

In some embodiments, a fluoropolymer component is combined with apolymer component. In some embodiments the fluoropolymer and the polymerare not miscible. In some embodiments, the fluoropolymer and the polymerare miscible.

According to other embodiments, a microfluidic device includes a firstcomponent including a polymer having a first curable functional groupand a second component including a polymer having a second curablefunctional group. In some embodiments, the first curable functionalgroup includes a photocurable functional group and the second curablefunctional group includes a thermalcurable functional group. In someembodiments, the first curable functional includes a first photocurablefunctional group and the second curable functional group includes asecond photocurable functional group and the first photocurablefunctional group is curable at a first wavelength and the secondphotocurable functional group is curable at a second wavelength. In someembodiments, the first curable functional group includes a firstthermalcurable functional group and the second curable functional groupincludes a second thermalcurable functional group and the firstthermalcurable functional group is curable at a first temperature andthe second thermalcurable functional group is curable at a secondtemperature. According to some embodiments, the first curable functionalgroup includes a first thermalcurable functional group and the secondcurable functional group includes a second thermalcurable functionalgroup and the first thermalcurable functional group is curable at afirst elapsed time and the second thermalcurable functional group iscurable at a second elapsed time.

In some embodiments, the photocurable functional group includes aphotocurable diurethane methacrylate and the thermalcurable functionalgroup includes a diisocyanate. In other embodiments, the photocurablefunctional group includes a photocurable diurethane methacrylate and thethermalcurable functional group includes a diepoxy. In some embodiments,the photocurable functional group includes a photocurable diurethanemethacrylate and the thermalcurable functional group includes a diamine.In some embodiments, the first photocurable functional group includes aphotocurable diurethane methacrylate and the second photocurablefunctional group includes a photocurable diepoxy.

According to some embodiments, the composition further includes apolymer having a third curable functional group. In some embodiments,the first curable functional group includes a photocurable diurethanemethacrylate, the second curable functional group includes athermalcurable diisocyanate, and the third curable functional groupincludes a thermalcurable triol. In some embodiments, the first curablefunctional group includes a photocurable diurethane methacrylate, thesecond curable functional group includes a thermalcurable diisocyanate,and the third curable functional group includes a thermalcurable tetrol.In some embodiments, the first curable functional group includes aphotocurable diurethane methacrylate, the second curable functionalgroup includes a thermalcurable diepoxy, and the third curablefunctional group includes a thermalcurable diamine.

According to some embodiments of the present invention, a microfluidicdevice includes a membrane. The membrane comprises a first componentincluding a polymer having a first curable functional group and a secondcomponent including a polymer having a second curable functional group.In some embodiments, the first curable functional group includes aphotocurable functional group and the second curable functional groupincludes a thermalcurable functional group. In some embodiments, thefirst curable functional group includes a first photocurable functionalgroup and the second curable functional group includes a secondphotocurable functional group and the first photocurable functionalgroup is curable at a first wavelength and the second photocurablefunctional group is curable at a second wavelength. In otherembodiments, the first curable functional group includes a firstthermalcurable functional group and the second curable functional groupincludes a second thermalcurable functional group and the firstthermalcurable functional group is curable at a first temperature andthe second thermalcurable functional group is curable at a secondtemperature. In yet other embodiments, the first curable functionalgroup includes a first thermalcurable functional group and the secondcurable functional group includes a second thermalcurable functionalgroup and the first thermalcurable functional group is curable at afirst elapsed time and the second thermalcurable functional group iscurable at a second elapsed time.

According to some embodiments, a microfluidic device includes expandedpolytetrafluoroethylene having perfluoropolyether cured in a pore of theexpanded polytetrafluoroethylene. In some embodiments, theperfluoropolyether includes a curable functional group. According tosome embodiments, the curable functional group includes a photocurablefunctional group, such as a photocurable diurethane methacrylate or aphotocurable diepoxy. In other embodiments, the curable function groupincludes a thermalcurable functional group, such as a thermalcurablediisocyanate, diepoxy, diamine, triol, tetrol, or combinations thereof.In other embodiments, the perfluoropolyether includes aperfluoropolyether distyrene.

According to some embodiments, a method of making a microfluidic deviceincludes providing a surface composed at least partially of expandedpolytetrafluoroethylene, wetting the expanded polytetrafluoroethylenewith a curable perfluoropolyether such that the perfluoropolyetherenters pores of the expanded polytetrafluoroethylene and curing theperfluoropolyether. In some embodiments, the curable perfluoropolyetherincludes a curable functional group and a second curable functionalgroup. According to some embodiments, the curable functional groupincludes a photocurable functional group or a thermalcurable functionalgroup. In other embodiments, the curable perfluoropolyether includes aphotocurable functional group and a thermalcurable functional group.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1C are a series of schematic end views depicting the formationof a patterned layer of polymeric material in accordance with thepresently disclosed subject matter.

FIGS. 2A-2D are a series of schematic end views depicting the formationof a device comprising two patterned layers of a polymeric material inaccordance with the presently disclosed subject matter.

FIGS. 3A-3C are schematic representations of an embodiment of thepresently disclosed method for adhering a functional device to a treatedsubstrate.

FIGS. 4A-4C are schematic representations of an embodiment of thepresently disclosed method for fabricating a multilayer device.

FIGS. 5A and 5B are schematic representations of an embodiment of thepresently disclosed method for functionalizing the interior surface of achannel.

FIG. 5A is a schematic representation of an embodiment of the presentlydisclosed method for functionalizing the interior surface of a channel.

FIG. 5B is a schematic representation of an embodiment of the presentlydisclosed method for functionalizing a surface of a device.

FIGS. 6A-6D are schematic representations of an embodiment of thepresently disclosed method for fabricating a microstructure using adegradable and/or selectively soluble material.

FIGS. 7A-7C are schematic representations of an embodiment of thepresently disclosed method for fabricating complex structures in adevice using degradable and/or selectively soluble materials.

FIG. 8 is a schematic plan view of a device in accordance with thepresently disclosed subject matter.

FIG. 9 is a schematic of an integrated microfluidic system forbiopolymer synthesis.

FIG. 10 is a schematic view of a system for flowing a solution orconducting a chemical reaction in a microfluidic device in accordancewith the presently disclosed subject matter.

FIGS. 11 a-11 e illustrate a process for fabricating a device accordingto an embodiment of the present invention.

FIGS. 12A-12B are photomicrographs of an air-actuated pneumatic valve ina presently disclosed PFPE microfluidic device actuated at a pressure ofabout 45 psi. FIG. 12A is a photomicrograph of an open valve and FIG.12B is a photomicrograph of a valve closed at about 45 psi.

FIG. 13 shows fabrication of a device from materials and methods of anembodiment of the present invention.

FIG. 14 shows a system for patching a disrupted component usingmaterials and methods of an embodiment of the present invention.

FIG. 15 shows molding and reconstruction of a molded object according toan embodiment of the present invention.

FIGS. 16A-16C show a device with a lumen according to an embodiment ofthe present invention.

FIG. 17 shows a device being formed through a master template patterningsequence according to an embodiment of the present invention.

FIG. 18 shows a sacrificial layer fabrication method and deviceaccording to an embodiment of the present invention.

DETAILED DESCRIPTION

The presently disclosed subject matter will now be described more fullyhereinafter with reference to the accompanying Drawings and Examples, inwhich representative embodiments are shown. The presently disclosedsubject matter can, however, be embodied in different forms and shouldnot be construed as limited to the embodiments set forth herein. Rather,these embodiments are provided so that this disclosure will be thoroughand complete, and will fully convey the scope of the embodiments tothose skilled in the art.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this presently described subject matter belongs. Allpublications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety.

Throughout the specification and claims, a given chemical formula orname shall encompass all optical and stereoisomers, as well as racemicmixtures where such isomers and mixtures exist.

I. Definitions

As used herein, the term “microfluidic device” generally refers to adevice through which materials, particularly fluid borne materials, suchas liquids, can be transported, in some embodiments on a micro-scale,and in some embodiments on a nano-scale. Thus, the microfluidic devicesdescribed by the presently disclosed subject matter can includemicroscale features, nanoscale features, and/or combinations thereof.

Accordingly, a microfluidic device typically includes structural orfunctional features dimensioned on the order of a millimeter-scale orless, which are capable of manipulating a fluid at a flow rate on theorder of a microliter/min or less. Typically, such features include, butare not limited to channels, fluid reservoirs, reaction chambers, mixingchambers, and separation regions. In some examples, the channels includeat least one cross-sectional dimension that is in a range of from about0.1 μm to about 500 μm. The use of dimensions on this order allows theincorporation of a greater number of channels in a smaller area, andutilizes smaller volumes of fluids.

A microfluidic device can exist alone or can be a part of a microfluidicsystem which, for example and without limitation, can include: pumps forintroducing fluids, e.g., samples, reagents, buffers and the like, intothe system and/or through the system; detection equipment or systems;reagent, product or data storage systems; and control systems forcontrolling fluid transport and/or direction within the device,monitoring and controlling environmental conditions to which fluids inthe device are subjected, e.g., temperature, current, and the like.

As used herein, the term “device” includes, but is not limited to, amicrofluidic device, a microtiter plate, tubing, hose, medical implantdevices, surgical devices, patches, orthopedic implants, medical devicecomponents, tools, and the like.

As used herein, the terms “channel,” “microscale channel,” and“microfluidic channel” are used interchangeably and can mean a recess orcavity formed in a material by imparting a pattern from a patternedsubstrate into a material or by any suitable material removingtechnique, or can mean a recess or cavity in combination with anysuitable fluid-conducting structure mounted in the recess or cavity,such as a tube, capillary, or the like.

As used herein, the terms “flow channel” and “control channel” are usedinterchangeably and can mean a channel in a microfluidic device in whicha material, such as a fluid, e.g., a gas or a liquid, can flow through.More particularly, the term “flow channel” refers to a channel in whicha material of interest, e.g., a solvent or a chemical reagent, can flowthrough. Further, the term “control channel” refers to a flow channel inwhich a material, such as a fluid, e.g., a gas or a liquid, can flowthrough in such a way to actuate a valve or pump. More particularly,such a channel is filled with a gas or fluid that does not permeate thematerial of the microfluidic device, medical device, or medical implant.An example of such a gas includes sulfur hexafluoride. Examples of suchliquids include mineral oil, silicon oil, propylene glycol, and ethyleneglycol. Said another way, in some embodiments, the presently disclosed“flow channels” and/or “control channels” are impermeable to the fluid,e.g., a gas or a liquid as described immediately hereinabove, disposedand/or flowing therein.

As used herein, the term “valve” unless otherwise indicated refers to aconfiguration in which two channels are separated by an elastomericsegment, e.g., a PFPE segment that can be deflected into or retractedfrom one of the channels, e.g., a flow channel, in response to anactuation force applied to the other channel, e.g., a control channel.The term “valve” also includes one-way valves, which include channelsseparated by a bead. “Valve” can also mean a synthetic or naturalbiologic valve such as, for example, a vascular valve, heart valve, orthe like.

As used herein, the term “pattern” can mean a channel or a microfluidicchannel or an integrated network of microfluidic channels, which, insome embodiments, can intersect at predetermined points. A pattern alsocan include one or more of a micro- or nano-scale fluid reservoir, amicro- or nano-scale reaction chamber, a micro- or nano-scale mixingchamber, a micro- or nano-scale separation region, a surface texture, apattern on a surface that can include micro and/or nano recesses and/orprojections. The surface pattern can be regular or irregular.

As used herein, the term “intersect” can mean to meet at a point, tomeet at a point and cut through or across, or to meet at a point andoverlap. More particularly, as used herein, the term “intersect”describes an embodiment wherein two channels meet at a point, meet at apoint and cut through or across one another, or meet at a point andoverlap one another. Accordingly, in some embodiments, two channels canintersect, i.e., meet at a point or meet at a point and cut through oneanother, and be in fluid communication with one another. In someembodiments, two channels can intersect, i.e., meet at a point andoverlap one another, and not be in fluid communication with one another,as is the case when a flow channel and a control channel intersect.

As used herein, the term “communicate” (e.g., a first component“communicates with” or “is in communication with” a second component)and grammatical variations thereof are used to indicate a structural,functional, mechanical, electrical, optical, or fluidic relationship, orany combination thereof, between two or more components or elements. Assuch, the fact that one component is said to communicate with a secondcomponent is not intended to exclude the possibility that additionalcomponents can be present between, and/or operatively associated orengaged with, the first and second components.

In referring to the use of a microfluidic device for handling thecontainment or movement of fluid, the terms “in”, “on”, “into”, “onto”,“through”, and “across” the device generally have equivalent meanings.

As used herein, the term “monolithic” refers to a structure having oracting as a single, uniform structure.

As used herein, the term “non-biological organic materials” refers toorganic materials, i.e., those compounds having covalent carbon-carbonbonds, other than biological materials. As used herein, the term“biological materials” includes nucleic acid polymers (e.g., DNA, RNA)amino acid polymers (e.g., enzymes, proteins, and the like) and smallorganic compounds (e.g., steroids, hormones) wherein the small organiccompounds have biological activity, especially biological activity forhumans or commercially significant animals, such as pets and livestock,and where the small organic compounds are used primarily for therapeuticor diagnostic purposes. While biological materials are of interest withrespect to pharmaceutical and biotechnological applications, a largenumber of applications involve chemical processes that are enhanced byother than biological materials, i.e., non-biological organic materials.

As used herein, the term “partial cure” refers to a process wherein lessthan about 100% of the polymerizable groups are reacted. Thus, the term“partially-cured material” refers to a material which has undergone apartial cure process.

As used herein, the term “full cure” refers to a process wherein about100% of the polymerizable groups are reacted. Thus, the term“fully-cured material” refers to a material which has undergone a fullcure process.

As used herein, the term “photocured” refers to the reaction ofpolymerizable groups whereby the reaction can be triggered by actinicradiation, such as UV light. In this application UV-cured can be asynonym for photocured.

As used herein, the term “thermal cure” or “thermally cured” refers tothe reaction of polymerizable groups, whereby the reaction can betriggered by heating the material beyond a threshold.

Following long-standing patent law convention, the terms “a”, “an”, and“the” refer to “one or more” when used in this application, includingthe claims. Thus, for example, reference to “a microfluidic channel”includes a plurality of such microfluidic channels, and so forth.

II. Materials

In certain embodiments, the presently disclosed subject matter broadlydescribes and employs solvent resistant, low surface energy polymericmaterials. According to some embodiments the low surface energypolymeric materials include, but are not limited to perfluoropolyether(PFPE), poly(dimethylsiloxane) (PDMS), poly(tetramethylene oxide),poly(ethylene oxide), poly(oxetanes), polyisoprene, polybutadiene,fluoroolefin-based fluoroelastomers, and the like. An example of castinga device with such materials includes casting liquid PFPE precursormaterials onto a patterned substrate and then curing the liquid PFPEprecursor materials to generate a patterned layer of functional PFPEmaterial, which can be used to form a device, such as a medical deviceor a microfluidic device. For simplification purposes, most of thedescription will focus on PFPE materials, however, it should beappreciated that other such polymers, such as those recited above, canbe equally applied to the methods, materials, and devices of the presentinvention.

Representative solvent resistant elastomer-based materials include butare not limited to fluorinated elastomer-based materials. As usedherein, the term “solvent resistant” refers to a material, such as anelastomeric material that neither swells nor dissolves in commonhydrocarbon-based organic solvents or acidic or basic aqueous solutions.Representative fluorinated elastomer-based materials include but are notlimited to perfluoropolyether (PFPE)-based materials.

In certain embodiments, functional liquid PFPE materials exhibitdesirable properties for use in a microfluidic and/or medical device.For example, functional PFPE materials typically have a low surfaceenergy, are non-toxic, UV and visible light transparent, highly gaspermeable; cure into a tough, durable, highly fluorinated elastomeric orglassy materials with excellent release properties, resistant toswelling, solvent resistant, biocompatible, combinations thereof, andthe like. The properties of these materials can be tuned over a widerange through the judicious choice of additives, fillers, reactiveco-monomers, and functionalization agents, examples of which aredescribed further herein.

Such properties that are desirable to modify, include, but are notlimited to, modulus, tear strength, surface energy, permeability,functionality, mode of cure, solubility, toughness, hardness,elasticity, swelling characteristics, combinations thereof, and thelike. Some examples of methods of adjusting mechanical and or chemicalproperties of the finished material includes, but are not limited to,shortening the molecular weight between cross-links to increase themodulus of the material, adding monomers that form polymers of high Tgto increase the modulus of the material, adding charged monomer orspecies to the material to increase the surface energy or wettability ofthe material, combinations thereof, and the like. In some embodiments,the materials of the present invention have a low surface energy,wherein low is defined as about 30 dynes/cm. According to oneembodiment, the surface energy is below about 30 dynes/cm. According toanother embodiment the surface energy is less than about 20 dynes/cm.According to a more preferred embodiment, the surface energy is lessthan about 18 dynes/cm. According to yet another embodiment the surfaceenergy is less than about 15 dynes/cm. According to still a furtherembodiment the surface energy is less than about 12 dynes/cm. Accordingto further embodiments the surface energy is less than about 10dynes/cm.

In some embodiments, the fluorinated polymers of the present invention,such as but not limited to PFPE, swell less than about a 15 percentvolume change when subjected to a hydrocarbon solvent. In alternativeembodiments, the hydrocarbon solvent can be tetrahydrofuran, toluene,dichloromethane, hexanes, chloroform, isopropyl alcohol, cyclohexane,methyl ethyl ketone, acetone, combinations thereof, and the like. Inalternative embodiments, the fluoropolymer of the present invention canbe altered, according to embodiments and methods herein, to decrease theswelling of the fluoropolymer to such solvents to less than about 10percent by weight. In further embodiments, the swelling of thefluoropolymer in response to the solvents listed herein is less thanabout an 8 percent weight change. In yet further embodiments, thecomposition and organization of the fluoropolymer can be arranged, bymethods disclosed herein such that the swelling of the fluoropolymer inresponse to the solvents listed herein is less than about a 5 percentweight change. Further solvents that can be applied to the materialsdisclosed herein can be found in Lee J. N. et al., Solvent Compatibilityof Poly(dimethylsiloxane) Based Microfluidic Devices, Anal. Chem. 75,6544-6554 (2003), which is incorporated herein by reference in itsentirety including all references cited therein. The non-swelling natureand easy release properties of the presently disclosed PFPE materialsallow for the fabrication of microfluidic devices.

According to some embodiments, the fluoropolymer of the presentinvention has a carbon to fluorine ratio of about 1:2. In someembodiments, the carbon to fluorine ratio is about 1:1. In alternativeembodiments, the carbon to fluorine ratio is about 2:1. According to yetother embodiments the carbon to fluorine ratio is between about 1:1 toabout 2:1. According to still further embodiments, the carbon tofluorine ratio is between about 1:1 to about 1:2.

According to an aspect of the present invention, the materials describedherein the molecular weight of the polymer can be selected or modified,for example by chain extension, mixing with other components, increasingor decreasing functional groups, and the like, to yield a resultingmaterial with predetermined mechanical properties, such as modulus,percent elongation at break, toughness, shear, combinations thereof, andthe like. In some embodiments, the modulus of the fluoropolymermaterials described herein can be decreased by increasing the molecularweight of the fluoropolymer. In one embodiment, the molecular weight ofthe fluoropolymer is about 16000 and the modulus is about 800 kPa.According to other embodiments, the modulus of the material can be about500 kPa. According to other embodiments, the modulus of the material canbe about 400 kPa. According to other embodiments, the modulus of thematerial can be about 200 kPa. According to other embodiments, themodulus of the material can be about 100 kPa. According to otherembodiments, the modulus of the material can be between about 100 kPaand about 5 megaPa.

In some embodiments, the percent elongation at break of the materialincreases as the molecular weight of the material increases. Accordingto some embodiments, the percent elongation at break of thefluoropolymers of the present invention is about 200 percent and themolecular weight is about 16000. According to an embodiment, the percentelongation at break of the fluoropolymer is about 300 percent. Accordingto an embodiment, the percent elongation at break of the fluoropolymeris about 250 percent. According to an embodiment, the percent elongationat break of the fluoropolymer is about 200 percent. According to anembodiment, the percent elongation at break of the fluoropolymer isabout 100 percent. According to some embodiments, the percent elongationat break of the fluoropolymer is between about 300 percent and about 100percent.

II.A. Perfluoropolyether Materials Prepared from a Liquid PFPE PrecursorMaterial Having a Viscosity Less than about 100 Centistokes

As would be recognized by one of ordinary skill in the art,perfluoropolyethers (PFPEs) have been in use for over 25 years for manyapplications. Commercial PFPE materials are made by polymerization ofperfluorinated monomers. The first member of this class was made by thecesium fluoride catalyzed polymerization of hexafluoropropene oxide(HFPO) yielding a series of branched polymers designated as KRYTOX®(DuPont, Wilmington, Del., United States of America). A similar polymeris produced by the UV catalyzed photo-oxidation of hexafluoropropene(FOMBLIN® Y) (Solvay Solexis, Brussels, Belgium). Further, a linearpolymer (FOMBLIN® Z) (Solvay) is prepared by a similar process, bututilizing tetrafluoroethylene. Finally, a fourth polymer (DEMNUM®)(Daikin Industries, Ltd., Osaka, Japan) is produced by polymerization oftetrafluorooxetane followed by direct fluorination. Structures for thesefluids are presented in Table I. Table II contains property data forsome members of the PFPE class of lubricants. Likewise, the physicalproperties of functional PFPEs are provided in Table III. In addition tothese commercially available PFPE fluids, a new series of structures arebeing prepared by direct fluorination technology. Representativestructures of these new PFPE materials appear in Table IV. Of theabovementioned PFPE fluids, only KRYTOX® and FOMBLIN® Z have beenextensively used in applications. See Jones, W. R., Jr., The Propertiesof Perfluoropolyethers Used for Space Applications, NASA TechnicalMemorandum 106275 (July 1993), which is incorporated herein by referencein its entirety. Accordingly, the use of such PFPE materials is providedin the presently disclosed subject matter.

TABLE I NAMES AND CHEMICAL STRUCTURES OF COMMERCIAL PFPE FLUIDS NAMEStructure DEMNUM ® C₃F₇O(CF₂CF₂CF₂O)_(x)C₂F₅ KRYTOX ®C₃F₇O[CF(CF₃)CF₂O]_(x)C₂F₅ FOMBLIN ® YC₃F₇O[CF(CF₃)CF₂O]_(x)(CF₂O)_(y)C₂F₅ FOMBLIN ® ZCF₃O(CF₂CF₂O)_(x)(CF₂O)_(y)CF₃

TABLE II PFPE PHYSICAL PROPERTIES Average Viscosity Vapor Molecular at20° C., Viscosity Pour Torr Pressure, Lubricant Weight (cSt) IndexPoint, ° C. 20° C. 100° C. FOMBLIN ® 9500 255 355 −66 2.9 × 10⁻¹² 1 ×10⁻⁸ Z-25 KRYTOX ® 3700 230 113 −40 1.5 × 10⁻⁶  3 × 10⁻⁴ 143AB KRYTOX ®6250 800 134 −35  2 × 10⁻⁸ 8 × 10⁻⁶ 143AC DEMNUM ® 8400 500 210 −53   1× 10⁻¹⁰ 1 × 10⁻⁷ S-200

TABLE III PFPE PHYSICAL PROPERTIES OF FUNCTIONAL PFPEs Average ViscosityMolecular at 20° C., Vapor Pressure, Torr Lubricant Weight (cSt) 20° C.100° C. FOMBLIN ® 2000 85 2.0 × 10⁻⁵ 2.0 × 10⁻⁵ Z-DOL 2000 FOMBLIN ®2500 76 1.0 × 10⁻⁷ 1.0 × 10⁻⁴ Z-DOL 2500 FOMBLIN ® 4000 100 1.0 × 10⁻⁸1.0 × 10⁻⁴ Z-DOL 4000 FOMBLIN ® 500 2000 5.0 × 10⁻⁷ 2.0 × 10⁻⁴ Z-TETROL

TABLE IV Names and Chemical Structures of Representative PFPE FluidsName Structure^(a) Perfluoropoly(methylene oxide) (PMO)CF₃O(CF₂O)_(x)CF₃ Perfluoropoly(ethylene oxide) (PEO)CF₃O(CF₂CF₂O)_(x)CF₃ Perfluoropoly(dioxolane) (DIOX)CF₃O(CF₂CF₂OCF₂O)_(x)CF₃ Perfluoropoly(trioxocane) (TRIOX)CF₃O[(CF₂CF₂O)₂CF₂O]_(x)CF₃ ^(a)wherein x is any integer.

In some embodiments, the perfluoropolyether precursor includespoly(tetrafluoroethylene oxide-co-difluoromethylene oxide)α,ω diol,which in some embodiments can be photocured to form one of aperfluoropolyether dimethacrylate and a perfluoropolyether distyreniccompound. A representative scheme for the synthesis and photocuring of afunctionalized perfluoropolyether is provided in Scheme 1.

II.B. Perfluoropolyether Materials Prepared from a Liquid PFPE PrecursorMaterial Having a Viscosity Greater than about 100 Centistokes

The methods provided herein below for promoting and/or increasingadhesion between a layer of a PFPE material and another material and/ora substrate and for adding a chemical functionality to a surface includea PFPE material having a characteristic selected from the groupconsisting of a viscosity greater than about 100 centistokes (cSt) and aviscosity less than about 100 cSt, provided that the liquid PFPEprecursor material having a viscosity less than 100 cSt is not afree-radically photocurable PFPE material. As provided herein, theviscosity of a liquid PFPE precursor material refers to the viscosity ofthat material prior to functionalization, e.g., functionalization with amethacrylate or a styrenic group.

Thus, in some embodiments, PFPE material is prepared from a liquid PFPEprecursor material having a viscosity greater than about 100 centistokes(cSt). In some embodiments, the liquid PFPE precursor is end-capped witha polymerizable group. In some embodiments, the polymerizable group isselected from the group consisting of an acrylate, a methacrylate, anepoxy, an amino, a carboxylic, an anhydride, a maleimide, an isocyanato,an olefinic, and a styrenic group.

In some embodiments, the perfluoropolyether material includes a backbonestructure selected from the group consisting of:

wherein X is present or absent, and when present includes an endcappinggroup, and n is an integer from 1 to 100.

In some embodiments, the PFPE liquid precursor is synthesized fromhexafluoropropylene oxide or tetrafluoro ethylene oxide as shown inScheme 2.

In some embodiments, the liquid PFPE precursor is synthesized fromhexafluoropropylene oxide or tetrafluoro ethylene oxide as shown inScheme 3.

In some embodiments the liquid PFPE precursor includes a chain extendedmaterial such that two or more chains are linked together before addingpolymerizablable groups. Accordingly, in some embodiments, a “linkergroup” joins two chains to one molecule. In some embodiments, as shownin Scheme 4, the linker group joins three or more chains.

In some embodiments, X is selected from the group consisting of anisocyanate, an acid chloride, an epoxy, and a halogen. In someembodiments, R is selected from the group consisting of an acrylate, amethacrylate, a styrene, an epoxy, a carboxylic, an anhydride, amaleimide, an isocyanate, an olefinic, and an amine. In someembodiments, the circle represents any multifunctional molecule. In someembodiments, the multifunctional molecule includes a cyclic molecule.PFPE refers to any PFPE material provided hereinabove.

In some embodiments, the liquid PFPE precursor includes a hyperbranchedpolymer as provided in Scheme 5, wherein PFPE refers to any PFPEmaterial provided hereinabove.

In some embodiments, the liquid PFPE material includes anend-functionalized material selected from the group consisting of:

In some embodiments the PFPE liquid precursor is encapped with an epoxymoiety that can be photocured using a photoacid generator. Photoacidgenerators suitable for use in the presently disclosed subject matterinclude, but are not limited to: bis(4-tert-butylphenyl)iodoniump-toluenesulfonate, bis(4-tert-butylphenyl)iodonium triflate,(4-bromo-phenyl)diphenylsulfonium triflate,(tert-butoxycarbonylmethoxynaphthyl)-diphenylsulfonium triflate,(tert-butoxycarbonylmethoxyphenyl)diphenyl-sulfonium triflate,(4-tert-butylphenyl)diphenylsulfonium triflate,(4-chlorophenyl)diphenylsulfonium triflate,diphenyliodonium-9,10-dimethoxy-anthracene-2-sulfonate, diphenyliodoniumhexafluorophosphate, diphenyliodonium nitrate, diphenyliodoniumperfluoro-1-butanesulfonate, diphenyliodonium p-toluene-sulfonate,diphenyliodonium triflate, (4-fluorophenyl)diphenylsulfonium triflate,N-hydroxynaphthalimide triflate,N-hydroxy-5-norbornene-2,3-dicarboximide perfluoro-1-butanesulfonate,N-hydroxyphthalimide triflate,[4-[(2-hydroxytetradecyl)oxy]phenyl]-phenyliodoniumhexafluoroantimonate, (4-iodophenyl)diphenylsulfonium triflate,(4-methoxyphenyl)diphenylsulfonium triflate,2-(4-methoxystyryl)-4,6-bis(trichloromethyl)-1,3,5-triazine,(4-methylphenyl)di-phenylsulfonium triflate, (4-methylthiophenyl)methylphenyl sulfonium triflate, 2-naphthyl diphenylsulfonium triflate,(4-phenoxy-phenyl)diphenylsulfonium triflate,(4-phenylthiophenyl)diphenylsulfonium triflate, thiobis(triphenylsulfonium hexafluorophosphate), triarylsulfonium hexafluoroantimonatesalts, triarylsulfonium hexafluorophosphate salts, triphenylsulfoniumperfluoro-1-butanesulfonate, triphenylsulfonium triflate,tris(4-tert-butylphenyl)sulfonium perfluoro-1-butanesulfonate, andtris(4-tert-butylphenyl)sulfonium triflate.

In some embodiments the liquid PFPE precursor cures into a highly UVand/or highly visible light transparent elastomer. In some embodimentsthe liquid PFPE precursor cures into an elastomer that is highlypermeable to oxygen, carbon dioxide, and nitrogen, a property that canfacilitate maintaining the viability of biological fluids/cells disposedtherein. In some embodiments, additives are added or layers are createdto enhance the barrier properties of the device to molecules, such asoxygen, carbon dioxide, nitrogen, dyes, reagents, and the like.

In some embodiments, the material suitable for use with the presentlydisclosed subject matter includes a silicone material having afluoroalkyl functionalized polydimethylsiloxane (PDMS) having thefollowing structure:

wherein:

R is selected from the group consisting of an acrylate, a methacrylate,and a vinyl group;

R_(f) includes a fluoroalkyl chain; and

n is an integer from 1 to 100,000.

In some embodiments, the material suitable for use with the presentlydisclosed subject matter includes a styrenic material having afluorinated styrene monomer selected from the group consisting of:

wherein R_(f) includes a fluoroalkyl chain.

In some embodiments, the material suitable for use with the presentlydisclosed subject matter includes an acrylate material having afluorinated acrylate or a fluorinated methacrylate having the followingstructure:

wherein:

R is selected from the group consisting of H, alkyl, substituted alkyl,aryl, and substituted aryl; and

R_(f) includes a fluoroalkyl chain with a —CH₂— or a —CH₂—CH₂— spacerbetween a perfluoroalkyl chain and the ester linkage. In someembodiments, the perfluoroalkyl group has hydrogen substituents.

In some embodiments, the material suitable for use with the presentlydisclosed subject matter includes a triazine fluoropolymer having afluorinated monomer.

In some embodiments, the fluorinated monomer or fluorinated oligomerthat can be polymerized or crosslinked by a metathesis polymerizationreaction includes a functionalized olefin. In some embodiments, thefunctionalized olefin includes a functionalized cyclic olefin.

According to an alternative embodiment, the PFPE material includes aurethane block as described and shown in the following structuresprovided in Scheme 6:

According to an embodiment of the present invention, PFPE urethanetetrafunctional methacrylate materials such as the above described canbe used as the materials and methods of the present invention or can beused in combination with other materials and methods described herein,as will be appreciated by one of ordinary skill in the art.

According to some embodiments, urethane systems include materials withthe following structures:

According to this scheme (Scheme 7), part A is a UV curable precursorand parts B and C make up a thermally curable component of the urethanesystem. The fourth precursor component, part D, is an end-cappedprecursor, (e.g., styrene end-capped liquid precursor). According tosome embodiments, part D reacts with latent methacrylate, acrylate, orstyrene groups contained in a base material, thereby adding chemicalcompatibility or a surface passivation to the base material andincreasing the functionality of the base material.

II.C. Fluoroolefin-based Materials

Further, in some embodiments, the materials used herein are selectedfrom highly fluorinated fluoroelastomers, e.g., fluoroelastomers havingat least fifty-eight weight percent fluorine, as described in U.S. Pat.No. 6,512,063 to Tang, which is incorporated herein by reference in itsentirety. Such fluoroelastomers can be partially fluorinated orperfluorinated and can contain between 25 to 70 weight percent, based onthe weight of the fluoroelastomer, of copolymerized units of a firstmonomer, e.g., vinylidene fluoride (VF₂) or tetrafluoroethylene (TFE).The remaining units of the fluoroelastomers include one or moreadditional copolymerized monomers, which are different from the firstmonomer, and are selected from the group consisting offluorine-containing olefins, fluorine containing vinyl ethers,hydrocarbon olefins, and combinations thereof.

These fluoroelastomers include VITON® (DuPont Dow Elastomers,Wilmington, Del., United States of America) and Kel-F type polymers, asdescribed for microfluidic applications in U.S. Pat. No. 6,408,878 toUnger et al. These commercially available polymers, however, have Mooneyviscosities ranging from about 40 to 65 (ML 1+10 at 121° C.) giving thema tacky, gum-like viscosity. When cured, they become a stiff, opaquesolid. As currently available, VITON® and Kel-F have limited utility formicro-scale molding. Curable species of similar compositions, but havinglower viscosity and greater optical clarity, is needed in the art forthe applications described herein. A lower viscosity (e.g., 2 to 32 (ML1+10 at 121° C.)) or more preferably as low as 80 to 2000 cSt at 20° C.,composition yields a pourable liquid with a more efficient cure.

More particularly, the fluorine-containing olefins include, but are notlimited to, vinylidine fluoride, hexafluoropropylene (HFP),tetrafluoroethylene (TFE), 1,2,3,3,3-pentafluoropropene (1-HPFP),chlorotrifluoroethylene (CTFE) and vinyl fluoride.

The fluorine-containing vinyl ethers include, but are not limited toperfluoro(alkyl vinyl)ethers (PAVEs). More particularly, perfluoro(alkylvinyl)ethers for use as monomers include perfluoro(alkyl vinyl)ethers ofthe following formula:CF₂═CFO(R_(f)O)_(n)(R_(f)O)_(m)R_(f)wherein each R_(f) is independently a linear or branched C₁-C₆perfluoroalkylene group, and m and n are each independently an integerfrom 0 to 10.

In some embodiments, the perfluoro(alkyl vinyl)ether includes a monomerof the following formula:CF₂═CFO(CF₂CFXO)_(n)R_(f)wherein X is F or CF₃, n is an integer from 0 to 5, and R_(f) is alinear or branched C₁-C₆ perfluoroalkylene group. In some embodiments, nis 0 or 1 and R_(f) includes 1 to 3 carbon atoms. Representativeexamples of such perfluoro(alkyl vinyl)ethers include perfluoro(methylvinyl)ether (PMVE) and perfluoro(propyl vinyl)ether (PPVE).

In some embodiments, the perfluoro(alkyl vinyl)ether includes a monomerof the following formula:CF₂═CFO[(CF₂)_(m)CF₂CFZO)_(n)R_(f)wherein R_(f) is a perfluoroalkyl group having 1-6 carbon atoms, m is aninteger from 0 or 1, n is an integer from 0 to 5, and Z is F or CF₃. Insome embodiments, R_(f) is C₃F₇, m is 0, and n is 1.

In some embodiments, the perfluoro(alkyl vinyl)ether monomers includecompounds of the formula:CF₂═CFO[(CF₂CF{CF₃}O)_(n)(CF₂CF₂CF₂O)_(m)(CF2)_(p)]C_(x)F_(2x+1)wherein m and n each integers independently from 0 to 10, p is aninteger from 0 to 3, and x is an integer from 1 to 5. In someembodiments, n is 0 or l, m is 0 or 1, and x is 1.

Other examples of useful perfluoro(alkyl vinyl ethers) include:CF₂═CFOCF₂CF(CF₃)O(CF₂O)_(m)C_(n)F_(2n+1)wherein n is an integer from 1 to 5, m is an integer from 1 to 3. Insome embodiments, n is 1.

In embodiments wherein copolymerized units of a perfluoro(alkylvinyl)ether (PAVE) are present in the presently describedfluoroelastomers, the PAVE content generally ranges from 25 to 75 weightpercent, based on the total weight of the fluoroelastomer. If the PAVEis perfluoro(methyl vinyl)ether (PMVE), then the fluoroelastomercontains between 30 and 55 wt. % copolymerized PMVE units.

Hydrocarbon olefins useful in the presently described fluoroelastomersinclude, but are not limited to ethylene (E) and propylene (P). Inembodiments wherein copolymerized units of a hydrocarbon olefin arepresent in the presently described fluoroelastomers, the hydrocarbonolefin content is generally 4 to 30 weight percent.

Further, the presently described fluoroelastomers can, in someembodiments, include units of one or more cure site monomers. Examplesof suitable cure site monomers include: i) bromine-containing olefins;ii) iodine-containing olefins; iii) bromine-containing vinyl ethers; iv)iodine-containing vinyl ethers; v) fluorine-containing olefins having anitrile group; vi) fluorine-containing vinyl ethers having a nitrilegroup; vii) 1,1,3,3,3-pentafluoropropene (2-HPFP); viii)perfluoro(2-phenoxypropyl vinyl)ether; and ix) non-conjugated dienes.

The brominated cure site monomers can contain other halogens, preferablyfluorine. Examples of brominated olefin cure site monomers areCF₂═CFOCF₂CF₂CF₂OCF₂CF₂Br; bromotrifluoroethylene;4-bromo-3,3,4,4-tetrafluorobutene-1 (BTFB); and others such as vinylbromide, 1-bromo-2,2-difluoroethylene; perfluoroallyl bromide;4-bromo-1,1,2-trifluorobutene-1;4-bromo-1,1,3,3,4,4-hexafluorobutene;4-bromo-3-chloro-1,1,3,4,4-penta-fluorobutene;6-bromo-5,5,6,6-tetrafluorohexene; 4-bromoperfluorobutene-1 and3,3-difluoroallyl bromide. Brominated vinyl ether cure site monomersinclude 2-bromo-perfluoroethyl perfluorovinyl ether and fluorinatedcompounds of the class CF₂Br—R_(f)—O—CF═CF₂ (wherein R_(f) is aperfluoroalkylene group), such as CF₂BrCF₂O—CF═CF₂, and fluorovinylethers of the class ROCF═CFBr or ROCBr═CF₂ (wherein R is a lower alkylgroup or fluoroalkyl group), such as CH₃OCF═CFBr or CF₃CH₂OCF═CFBr.

Suitable iodinated cure site monomers include iodinated olefins of theformula: CHR═CH-Z-CH₂CHR—I, wherein R is —H or —CH₃; Z is a C₁ to C₁₈(per)fluoroalkylene radical, linear or branched, optionally containingone or more ether oxygen atoms, or a (per)fluoropolyoxyalkylene radicalas disclosed in U.S. Pat. No. 5,674,959. Other examples of usefuliodinated cure site monomers are unsaturated ethers of the formula:I(CH₂CF₂CF₂)_(n)OCF═CF₂ and ICH₂CF₂O[CF(CF₃)CF₂O]_(n)CF═CF₂, and thelike, wherein n is an integer from 1 to 3, such as disclosed in U.S.Pat. No. 5,717,036. In addition, suitable iodinated cure site monomersincluding iodoethylene, 4-iodo-3,3,4,4-tetrafluorobutene-1 (ITFB);3-chloro-4-iodo-3,4,4-trifluorobutene;2-iodo-1,1,2,2-tetrafluoro-1-(vinyloxy)ethane;2-iodo-1-(perfluorovinyloxy)-1,1-2,2-tetrafluoroethylene;1,1,2,3,3,3-hexafluoro-2-iodo-1-(perfluorovinyloxy)-propane; 2-iodoethylvinyl ether; 3,3,4,5,5,5-hexafluoro-4-iodopentene; andiodotrifluoroethylene are disclosed in U.S. Pat. No. 4,694,045. Allyliodide and 2-iodo-perfluoroethyl perfluorovinyl ether also are usefulcure site monomers.

Useful nitrile-containing cure site monomers include those of theformulas shown below:CF₂═CF—O(CF₂)_(n)—CNwherein n is an integer from 2 to 12. In some embodiments, n is aninteger from 2 to 6.CF₂═CF—O[CF₂—CF(CF)—O]_(n)—CF₂—CF(CF₃)—CNwherein n is an integer from 0 to 4. In some embodiments, n is aninteger from 0 to 2.CF₂═CF—[OCF₂CF(CF₃)]—O—(CF₂)_(n)—CNwherein x is 1 or 2, and n is an integer from 1 to 4; andCF₂═CF—O—(CF₂)_(n)—O—CF(CF₃)—CNwherein n is an integer from 2 to 4. In some embodiments, the cure sitemonomers are perfluorinated polyethers having a nitrile group and atrifluorovinyl ether group.

In some embodiments, the cure site monomer is:CF₂═CFOCF₂CF(CF₃)OCF₂CF₂CNi.e., perfluoro(8-cyano-5-methyl-3,6-dioxa-1-octene) or 8-CNVE.

Examples of non-conjugated diene cure site monomers include, but are notlimited to 1,4-pentadiene; 1,5-hexadiene; 1,7-octadiene;3,3,4,4-tetrafluoro-1,5-hexadiene; and others, such as those disclosedin Canadian Patent No. 2,067,891 and European Patent No. 0784064A1. Asuitable triene is 8-methyl-4-ethylidene-1,7-octadiene.

In embodiments wherein the fluoroelastomer will be cured with peroxide,the cure site monomer is preferably selected from the group consistingof 4-bromo-3,3,4,4-tetrafluorobutene-1 (BTFB);4-iodo-3,3,4,4-tetrafluorobutene-1 (ITFB); allyl iodide;bromotrifluoroethylene and 8-CNVE. In embodiments wherein thefluoroelastomer will be cured with a polyol, 2-HPFP orperfluoro(2-phenoxypropyl vinyl)ether is the preferred cure sitemonomer. In embodiments wherein the fluoroelastomer will be cured with atetraamine, bis(aminophenol) or bis(thioaminophenol), 8-CNVE is thepreferred cure site monomer.

Units of cure site monomer, when present in the presently disclosedfluoroelastomers, are typically present at a level of 0.05-10 wt. %(based on the total weight of fluoroelastomer), preferably 0.05-5 wt. %and most preferably between 0.05 and 3 wt. %.

Fluoroelastomers which can be used in the presently disclosed subjectmatter include, but are not limited to, those having at least 58 wt. %fluorine and having copolymerized units of i) vinylidene fluoride andhexafluoropropylene; ii) vinylidene fluoride, hexafluoropropylene andtetrafluoroethylene; iii) vinylidene fluoride, hexafluoropropylene,tetrafluoroethylene and 4-bromo-3,3,4,4-tetrafluorobutene-1; iv)vinylidene fluoride, hexafluoropropylene, tetrafluoroethylene and4-iodo-3,3,4,4-tetrafluorobutene-1; v) vinylidene fluoride,perfluoro(methyl vinyl)ether, tetrafluoroethylene and4-bromo-3,3,4,4-tetrafluorobutene-1; vi) vinylidene fluoride,perfluoro(methyl vinyl)ether, tetrafluoroethylene and4-iodo-3,3,4,4-tetrafluorobutene-1; vii) vinylidene fluoride,perfluoro(methyl vinyl)ether, tetrafluoroethylene and1,1,3,3,3-pentafluoropropene; viii) tetrafluoroethylene,perfluoro(methyl vinyl)ether and ethylene; ix) tetrafluoroethylene,perfluoro(methyl vinyl)ether, ethylene and4-bromo-3,3,4,4-tetrafluorobutene-1; x) tetrafluoroethylene,perfluoro(methyl vinyl)ether, ethylene and4-iodo-3,3,4,4-tetrafluorobutene-1; xi) tetrafluoroethylene, propyleneand vinylidene fluoride; xii) tetrafluoroethylene and perfluoro(methylvinyl)ether; xiii) tetrafluoroethylene, perfluoro(methyl vinyl)ether andperfluoro(8-cyano-5-methyl-3,6-dioxa-1-octene); xiv)tetrafluoroethylene, perfluoro(methyl vinyl)ether and4-bromo-3,3,4,4-tetrafluorobutene-1; xv) tetrafluoroethylene,perfluoro(methyl vinyl)ether and 4-iodo-3,3,4,4-tetrafluorobutene-1; andxvi) tetrafluoroethylene, perfluoro(methyl vinyl)ether andperfluoro(2-phenoxypropyl vinyl)ether.

Additionally, iodine-containing endgroups, bromine-containing endgroupsor combinations thereof can optionally be present at one or both of thefluoroelastomer polymer chain ends as a result of the use of chaintransfer or molecular weight regulating agents during preparation of thefluoroelastomers. The amount of chain transfer agent, when employed, iscalculated to result in an iodine or bromine level in thefluoroelastomer in the range of 0.005-5 wt. %, preferably 0.05-3 wt. %.

Examples of chain transfer agents include iodine-containing compoundsthat result in incorporation of bound iodine at one or both ends of thepolymer molecules. Methylene iodide; 1,4-diiodoperfluoro-n-butane; and1,6-diiodo-3,3,4,4-tetrafluorohexane are representative of such agents.Other iodinated chain transfer agents include1,3-diiodoperfluoropropane; 1,6-diiodoperfluorohexane;1,3-diiodo-2-chloroperfluoropropane;1,2-di(iododifluoromethyl)perfluorocyclobutane; monoiodoperfluoroethane;monoiodoperfluorobutane; 2-iodo-1-hydroperfluoroethane, and the like.Also included are the cyano-iodine chain transfer agents disclosedEuropean Patent No. 0868447A1. Particularly preferred are diiodinatedchain transfer agents.

Examples of brominated chain transfer agents include1-bromo-2-iodoperfluoroethane; 1-bromo-3-iodoperfluoropropane;1-iodo-2-bromo-1,1-difluoroethane and others such as disclosed in U.S.Pat. No. 5,151,492.

Other chain transfer agents suitable for use include those disclosed inU.S. Pat. No. 3,707,529. Examples of such agents include isopropanol,diethylmalonate, ethyl acetate, carbon tetrachloride, acetone anddodecyl mercaptan.

II.D. Dual Photo-curable and Thermal-curable Materials

According to another embodiment, a material according to the inventionincludes one or more of a photo-curable constituent and athermal-curable constituent. In one embodiment, the photo-curableconstituent is independent from the thermal-curable constituent suchthat the material can undergo multiple cures. A material having theability to undergo multiple cures is useful, for example, in forminglayered devices or in connecting or attaching devices to other devicesor portions or components of devices to other portions or components ofdevices. For example, a liquid material having photocurable andthermal-curable constituents can undergo a first cure to form a firstdevice through, for example, a photocuring process or a thermal curingprocess. Then the photocured or thermal cured first device can beadhered to a second device of the same material or any material similarthereto that will thermally cure or photocure and bind to the materialof the first device. By positioning the first device and second deviceadjacent one another and subjecting the first and second devices to athermal curing or photocuring, whichever component that was notactivated on the first curing. Thereafter, either the thermal cureconstituents of the first device that were left un-activated by thephotocuring process or the photocure constituents of the first devicethat were left un-activated by the first thermal curing, will beactivated and bind the second device. In one embodiment, using thismethod and materials, the first and second devices become adheredtogether. It will be appreciated by one of ordinary skill in the artthat the order of curing processes is independently determined and athermal-curing could occur first followed by a photocuring or aphotocuring could occur first followed by a thermal curing.

According to yet another embodiment, multiple thermo-curableconstituents can be included in the material such that the material canbe subjected to multiple and independent thermal curing processes. Forexample, the multiple thermal-curable constituents can have differentactivation temperature ranges such that the material can undergo a firstthermal-cure at a first temperature range, a second thermal-cure at asecond temperature range, etc. Accordingly, in one embodiment, thematerial can be adhered to multiple other materials through differentthermal-cures, thereby, forming a multiple laminate layer device. Inalternative embodiments, the material can include different thermal-cureconstituents that react or are activated at different rates, thereby,presenting the user with the option to first cure the material at afirst rate, then subject the material to a second cure at a differentrate to adhere the first cured material to a second material forexample.

In another embodiment, the material of the present invention can includea dual photo-cure material which can include different constituents thatcan be cured at different wavelengths. For example, a first photo-cureconstituent can be a dimethacrylate that cures at a wavelength of about365 nm. The material can also include a second photo-cure constituent,such as, for example, a diepoxy material that is activated at anotherwavelength, such as for example 254 nm. In this manner, multiple layersof a device can be bonded and adhered to other substrates such as glass,silicon, other polymeric materials, laminates, combinations thereof, andthe like at different stages of fabrication of an overall device.

Examples of chemical groups which would be suitable end-capping agentsfor a UV curable component include: methacrylates, acrylates, styrenics,epoxides, cyclobutanes and other 2+2 cycloadditions, combinationsthereof, and the like. Examples of chemical group pairs which aresuitable to endcap a thermally curable component include: epoxy/amine,epoxy/hydroxyl, triol/diisocyanate, tetrol/diisocyanate, carboxylicacid/amine, carboxylic acid/hydroxyl, ester/amine, ester/hydroxyl,amine/anhydride, acid halide/hydroxyl, acid halide/amine, amine/halide,hydroxyl/halide, hydroxyl/chlorosilane, azide/acetylene and otherso-called “click chemistry” reactions, and metathesis reactionsinvolving the use of Grubb's-type catalysts, combinations thereof, andthe like. Examples of UV and thermal end-cap combinations, according tosome embodiments, include: UV diurethane methacrylate with thermal trioland diisocyanate components, UV diurethane methacrylate with thermaltetrol and diisocyanate components, UV diurethane methacrylate with UVdiepoxy, UV diurethane methacrylate with thermal diepoxy and diaminecomponents, UV diurethane methacrylate with thermal diisocyanate,combinations thereof, and the like.

The presently disclosed methods for the adhesion of multiple layers of adevice to one another or to a separate surface can be applied toPFPE-based materials, as well as a variety of other materials, includingPDMS and other liquid-like polymers. Examples of liquid-like polymericmaterials that are suitable for use in the presently disclosed adhesionmethods include, but are not limited to, PDMS, poly(tetramethyleneoxide), poly(ethylene oxide), poly(oxetanes), polyisoprene,polybutadiene, and fluoroolefin-based fluoroelastomers, such as thoseavailable under the registered trademarks VITON® AND KALREZ®.

Accordingly, in one embodiment, the presently disclosed methods can beused to adhere layers of different polymeric materials together to formdevices, such as microfluidic devices, medical device, surgical devices,tools, components of medical devices, implant materials, laminates, andthe like. For example, multiple PFPE and PDMS layers can be adheredtogether in a given microfluidic and/or medical device. In someembodiments, the different polymeric materials include a fluoropolymermixed with a polymer in the same device. According to some embodiments,the polymer is miscible with the fluoropolymer. According to otherembodiments, the polymer is immiscible with the fluoropolymer.

According to some embodiments, a device can be formed from a compositionof a fluoropolymer mixed with a polymer, wherein the fluoropolymerincludes a first functional group and the polymer includes a secondfunctional group. According to such embodiments, the functional groupscan be photocurable and/or thermalcurable functional groups as disclosedherein. In other embodiments, a device can be formed from a compositionhaving a first polymer and a second polymer, wherein the first polymerincludes a first functional group and the second polymer includes asecond functional group. According to such embodiments, the first andsecond functional groups can include functional groups disclosed herein,such as photocurable and/or thermalcurable functional groups. In yetother embodiments, a device can be formed from a composition thatincludes a first fluoropolymer and a second fluoropolymer, wherein thefirst fluoropolymer includes a first functional group and the secondfluoropolymer includes a second functional group. According to suchembodiments, the first and second functional groups can includefunctional groups disclosed herein, such as photocurable and/orthermalcurable functional groups. The compositions just described canundergo a first cure by activating the first curable functional group ofone of the components of the composition (i.e., the photocurablefunctional group), then, after multiple components of a device areformed (through photocuring of the materials), the components can bepositioned with respect to each other and treated to a second cure tothereby activate the second function group (i.e., thermalcurablefunctional group) and cure or laminate the multiple components together.

According to other embodiments, different components of a device can beformed from different materials, such as for example, a first componentof a device can be formed from a polymer material and a second componentof the device can be formed from a fluoropolymer material. According tosuch embodiments, both the polymer and the fluoropolymer can includedual cure functional groups, such as for example, photocurable andthermalcurable functional groups. Accordingly, the individual componentsof a device can be fabricated by subjecting the materials to a firstcure (i.e., a photocure) to fabricate the component, then the individualcomponents can be positioned with respect to each other to form thedesired device and subjected to a second cure (i.e., thermal cure) tobind the individual components and form the device.

II.E. Silicone Based Materials

According to alternate embodiments, novel silicone based materialsinclude one or more photocurable and thermal-curable components. In suchalternate embodiments, silicone based materials can include one or morephoto-curable and one or more thermal-curable components such that thesilicone based material has a dual curing capability as describedherein. Silicone based materials compatible with the present inventionare described herein and throughout the reference materials incorporatedby reference into this application.

II.F. Phosphazene-containing Polymers

According to some embodiments, devices and methods disclosed herein canbe formed with materials that include phosphazene-containing polymershaving the following structure. According to these embodiments, R, inthe structure below, can be a fluorine-containing alkyl chain. Examplesof such fluorine-containing alkyl chains can be found in Langmuir, 2005,21, 11604, the disclosure of which is incorporated herein by referencein its entirety. The devices disclosed in this application can be formedfrom phosphazene-containing polymers or from PFPE in combination withphosphazene-containing polymers.

II.G. Materials End-capped with an Aryl Trifluorovinyl Ether (TVE)

In some embodiments, devices and methods disclosed herein can be formedwith materials that include materials end-capped with one or more aryltrifluorovinyl ether (TVE) group, as shown in the structure below.Examples of materials end-capped with a TVE group can be found inMacromolecules, 2003, 36, 9000, which is incorporated herein byreference in its entirety. These structures react in a 2+2 addition atabout 150° C. to form perfluorocyclobutyl moieties. In some embodiments,Rf can be a fluoropolymer, for example a PFPE chain. In some embodimentsthree or more TVE groups are present on a 3-armed PFPE polymer such thatthe material crosslinks into a network.

II.H. Sodium Naphthalene Etchant

In some embodiments a sodium naphthalene etchant, such as commerciallyavailable Tetraetch™, is contacted with a layer of a device, such as afluoropolymer device disclosed herein. In other embodiments, a sodiumnaphthalene etchant is contacted with a layer of a PFPE-based device,such as a microfluidic device disclosed herein. According to suchembodiments, the etch reacts with C—F bonds in the polymer of the deviceforming functional groups along a surface of the device. In someembodiments, these functional groups can then be reacted with modalitieson other layers, on a silicon surface, on a glass surface, combinationsthereof, or the like, thereby forming an adhesive bond. In someembodiments, such adhesive bonds available on the surface of devicesdisclosed herein, such as microfluidic devices, can increase adhesionbetween two devices, layers of a device, combinations thereof, or thelike. Increasing the bonding strength between layers of a microfluidicdevice can increase the functionality of the device, for example, byincreasing the pressure range that channels of the device can beoperated within, increasing valve pressures, and the like.

II.I. Trifunctional PFPE Precursor

According to some embodiments, a trifunctional PFPE precursor can beused to fabricate devices disclosed herein, such as microfluidicdevices. In one embodiment, the trifunctional PFPE precursor disclosedherein can increase the functionality of an overall device by increasingthe number of functional groups that can be added to the material.Moreover, the trifunctional PFPE precursor can provide for increasedcross-linking capabilities of the material. According to suchembodiments, devices can be synthesized by the following reactionscheme.

In further embodiments, a trifunctional PFPE precursor for thefabrication of devices, such as for example microfluidic devicesdisclosed herein, is synthesized by the following reaction scheme.

II.J. Fluoroalkyliodide Precursors for Generating Fluoropolymers and/orPFPE's

In some embodiments, functional PFPEs or other fluoropolymers can begenerated using fluoroalkyliodide precursors. According to suchembodiments, such materials can be modified by insertion of ethylene andthen transformed into a host of common functionalities including but notlimited to: silanes, Gringard reagents, alcohols, cyano, thiol,epoxides, amines, and carboxylic acids.Rf-I+═→Rf-CH₂CH₂—I

II.K. Diepoxy Materials

According to some embodiments, one or more of the PFPE precursors usefulfor fabricating devices disclose herein, such as microfluidic devicesfor example, contains diepoxy materials. The diepoxy materials can besynthesized by reaction of PFPE diols with epichlorohydrin according tothe reaction scheme below.

II.L. Encapped PFPE Chains with Cycloaliphatic Epoxides

In some embodiments, PFPE chains can be encapped with cycloaliphaticepoxides moieties such as cyclohexane epoxides, cyclopentane epoxides,combinations thereof, or the like. In some embodiments, the PFPE diepoxyincludes a chain-extending material having the structure belowsynthesized by varying the ratio of diol to epichlorohydrin during thesynthesis. Examples of some synthesis procedures are described byTonelli et al. in Journal of Polymer Science: Part A: Polymer Chemistry1996, Vol 34, 3263, which is incorporated herein by reference in itsentirety. Utilizing this method, physical and chemical properties, suchas mechanical, optical, thermal, and the like, including elasticity,solvent resistance, translucency, toughness, adhesion, and the like, ofthe cured material can be tuned to predetermined requirements.

In further embodiments, the secondary alcohol formed in this reactioncan be used to attach further functional groups. An example of this isshown below whereby the secondary alcohol is reacted with2-isocyanatoethyl methacrylate to yield a material with species reactivetowards both free radical and cationic curing. In some embodiments,functional groups such as in this example can be utilized to bondsurfaces together, such as for example, layers of fluoropolymer, such asPFPE, material in a microfluidic device. In still further embodiments,moieties along the wall of a microfluidic chip such as biomolecules,proteins, charged species, catalysts, etc. can be attached through suchsecondary alcohol species.

II.M. PFPE Diepoxy Cured with Diamines

In some embodiments, PFPE diepoxy can be cured with traditionaldiamines, including but not limited to, 1,6 hexanediamine; isophoronediamine; 1,2 ethanediamine; combinations thereof; and the like.According to some embodiments the diepoxy can be cured with imidazolecompounds including those with the following or related structures whereR1, R2, and R3 can be a hydrogen atom or other alkyl substituents suchas methyl, ethyl, propyl, butyl, fluoroalkyl compounds, combinationsthereof, and the like. According to some embodiments, the imidazoleagent is added to the PFPE diepoxy in concentrations on the order ofbetween about 1-25 mol % in relation to the epoxy content. In someembodiments the PFPE diepoxy containing an imidazole catalyst is thethermal part of a two cure system, such as described elsewhere herein.

II.N. PFPE Cured with Photoacid Generators

In some embodiments, a PFPE diepoxy can be cured through the use ofphotoacid generators (PAGs). In some embodiments, the PAGs are dissolvedin the PFPE material in concentrations ranging from between about 1 toabout 5 mol % relative to epoxy groups and cured by exposure to UVlight. Specifically, for example, these photoacid generators can possesthe following structure (Rhodorsil™) 2074 (Rhodia, Inc):

In other embodiments, the photoacid generator can be, for example,Cyracure™ (Dow Corning) possessing the following structure:

II.O. PFPE Diol Containing a Poly(ethylene glycol)

In some embodiments, a commercially available PFPE diol containing a thenumber of poly(ethylene glycol) units, such as those commercially soldas ZDOL TX™ (Solvay Solexis) can be used as the material for fabricationof a device, such as a microfluidic device. In other embodiments, thecommercially available PFPE diol containing a given number ofpoly(ethylene glycol) units is used in combination with other materialsdisclosed herein. Such materials can be useful for dissolving the abovedescribed photoinitiators into the PFPE diepoxy and can also be helpfulin tuning mechanical properties of the material as the PFPE diolcontaining a poly(ethylene glycol) unit can react with propagating epoxyunits and can be incorporated into the final network.

II.P. PFPE Diols and/or Polyols Mixed with a PFPE Diepoxy

In further embodiments, commercially available PFPE diols and/orpolyols, shown below and commercially sold as ZDOL™ and Z-Tetrol™(Solvay Solexis) can be mixed with a PFPE diepoxy compound to tunemechanical properties by incorporating into the propagating epoxynetwork during curing.

II.Q. PFPE Epoxy-containing a PAG Blended with a Photoinitiator

In some embodiments, a PFPE epoxy-containing a PAG can be blended withbetween about 1 and about 5 mole % of a free radical photoinitiator suchas, for example, 2,2-dimethoxyacetophenone, 1-hydroxy cyclohexyl phenylketone, diethoxyacetophenone, combinations thereof, or the like. Thesematerials, when blended with a PAG, form reactive cationic species whichare the product of oxidation by the PAG when the free-radical initiatorsare activated with UV light, as partially described by Crivello et al.Macromolecules 2005, 38, 3584, which is incorporated herein by referencein its entirety. In some embodiments, such cationic species can becapable of initiating epoxy polymerization and/or curing. The use ofthis method allows the PFPE diepoxy to be cured at a variety ofdifferent wavelengths.

II.R. PFPE Diepoxy Containing a Photoacid Generator and Blended with aPFPE Diepoxy

In some embodiments, a PFPE diepoxy material containing a photoacidgenerator can be blended with a PFPE dimethacrylate material containinga free radical photoinitiator and possessing the following structure:

In one embodiment, the blended material includes a dual cure materialwhich can be cured at one wavelength, for example, curing thedimethacrylate at 365 nm, and then bonded to other layers of materialthrough activating the curing of the second diepoxy material at anotherwavelength, such as for example 254 nm. In this manner, multiple layersof patterned PFPE materials can be bonded and adhered to substrates suchas glass, silicon, other polymeric materials, combinations thereof, andthe like at different stages of fabrication of an overall device.

II.S. Other Materials

According to alternative embodiments, the following materials can beutilized alone, in connection with other materials disclosed herein, ormodified by other materials disclosed herein and applied to the methodsdisclosed herein to fabricate, in some embodiments devices disclosedherein. Moreover, end-groups disclosed herein and disclosed in U.S. Pat.Nos. 3,810,874; and 4,818,801, each of which is incorporated herein byreference including all references cited therein.

II.S.i. Diurethane Methacrylate

In some embodiments, the material is or includes diurethane methacrylatefabricated from the following reaction and having a modulus of about 4.0MPa and is UV curable with the following structure:

II.S.ii. Chain-extended Diurethane Methacrylate

In some embodiments, the material is or includes a chain extendeddiurethane methacrylate fabricated from the following reaction andhaving nomenclature: 2M-240, 2M-340, 2M-440; chain extension beforeend-capping which increases molecular weight between crosslinks; modulusof approximately 2.0 MPa; and is UV curable, having the followingstructure:

In some alternative embodiments, the chain extended diurethanemethacrylate has the following structure:

II.S.iii. Diisocyanate

In some embodiments, the material is or includes nomenclature: 2I-140;the material is typically one component of a two-component thermallycurable system; may be cured by itself through a moisture cure technique(disclosed herein); can be fabricated from the following reaction andhas the following structure:

In some alternative embodiments, the diisocyanate functionalizedperfluoropolyether has the following structure:

II.S.iv. Chain Extended Diisocyanate

In some embodiments, the material is or includes, nomenclature: 2I-240,2I-340, 2I-440 (depending on the degree of chain extension); where thematerial is one component of a two component thermally curable system;chain extended by linking several PFPE chains together; may be cured byitself through a moisture cure; can be fabricated from the followingreaction and includes the following structure:

II.S.v. Blocked Diisocyanate

In some embodiments, the material is or includes: nomenclature: 2I-140B;includes similar function to 2I-140 but not moisture sensitive (blockinggroup cleaves at high temp and reforms isocyanate); material can be onecomponent of a two component thermally curable system; can be fabricatedfrom the following reaction and includes the following structure:

II.S.vi. PFPE Three-armed Triol

In some embodiments, the material is or includes: nomenclature: 3A-115;is one component of a two-component thermally curable urethane system;includes the benefits of being highly miscible with other PFPEcompositions; can be fabricated from the following reaction and includesthe following structure:

II.S.vii. PFPE DiStyrene

In some embodiments, the material is or includes: nomenclature: 2S-140;is UV curable; highly chemically stable; is useful in making laminatecoatings with other compositions; can be fabricated from the followingreaction and includes the following structure:

II.S.viii. Diepoxy

In some embodiments, the material is or includes: nomenclature: 2E-115,2E-140 (depending on MW of the chain); can be UV cured; can be thermallycured by itself using imidazoles; can also be thermally cured in atwo-component diamine system; is highly chemically stable; can befabricated from the following reaction and includes the followingstructure:

II.S.ix. Diamine

In some embodiments, the material is or includes: nomenclature: 2A-115,2A-140 (depending on MW of the chain); can be thermally cured in atwo-component diamine system; has functionality of 6 (3 amines availableon each end); is highly chemically stable; can be fabricated from thefollowing reaction and includes the following structure:

II.S.x. Thermally Cured PU-Tetrol

In some embodiments, the material is or includes: nomenclature:2I-140+Z-tetrol; can be thermally cured in a two-component system, suchas for example mixed in a 2:1 molar ratio at about 100-about 130 degreesC.; forms tough, mechanically stable network; the cured network isslightly cloudy due to immiscibility of tetrol; and includes thefollowing:

II.S.xi. Thermally Cured PU-Triol

In some embodiments, the material is or includes: nomenclature:2I-140+3A-115; can be thermally cured in a two-component system, such asfor example mixed in a 3:2 molar ratio, at about 100-about 130 degreesC.; forms tough, mechanically stable network; where the cured network isclear and colorless; and includes the following:

II.S.xii Thermally Cured Epoxy

In some embodiments, the material is or includes: nomenclature:2E-115+2A-115; can be thermally cured in a two-component system, such asfor example mixed in a 3:1 molar ratio, at about 100-about 130 degreesC.; forms mechanically stable network; where the cured network is clearand colorless; has high chemical stability; and includes the following:

II.S.xiii. UV-Cured Epoxy

In some embodiments, the material is or includes: nomenclature:2E-115+ZDOL TX; is a UV curable composition; includes ZDOL TX used tosolubilize PAG; where the cured network is clear and yellow; has highchemical stability; and includes the following:

II.S.xiv. UV—Thermal Dual Cure

In some embodiments, the material is or includes: nomenclature: 2M-240,2I-140, Z-Tetrol; can be mixed in a 2:1 ratio (UV to thermal); formscloudy network (tetrol); has a high viscosity; forms a very strongadhesion; has very good mechanical properties; and includes thefollowing:

II.S.xv. Orthogonal Cure with Triol

In some embodiments, the material is or includes: nomenclature: 2M-240,2I-140, 3A-115; can be mixed in a 2:1 ratio (UV to thermal); forms clearand colorless network; has a high viscosity; forms very strong adhesion;includes very good mechanical properties; and includes the following:

II.S.xvi. UV Orthogonol System

In some embodiments, the material is or includes: nomenclature: 2M-240,2E-115, ZDOL-TX; can be mixed in a 1:1 ratio (epoxy to methacrylate);forms clear and yellow network; has strong adhesion properties; has goodmechanical properties; and includes the following:

II.S.xvii. UV with Epoxy Dual Cure

In some embodiments, the material is or includes: nomenclature: 2M-240,2E-115, DA-115; the material forms slightly yellow network; includes aratio (2:1 UV to thermal); has good mechanical properties; goodadhesion; is highly chemical stability; and includes the following:

II.S.xviii. Orthogonal with Diisocyanate

In some embodiments, the material is or includes: nomenclature: 2M-240,2I-140; is one component of a thermal cure system (isocyanate reactswith urethane linkage on urethane dimethacrylate); has good mechanicalproperties; forms a strong adhesion; cures to clear, slightly yellownetwork; and includes the following:

II.S.xix. ePTFE Combined with PFPE

In some embodiments, expanded poly(tetrafluoroethylene) (ePTFE) iscombined with PFPE. ePTFE is a microporous poly(tetrafluoroethylene)(PTFE) membrane formed by expanding PTFE at high temperatures. Becauseof the incredibly low surface energy of perfluoropolyether (PFPE)materials, as disclosed herein, the PFPE can be cast onto ePTFEmembranes and effectively wet the pores of the ePTFE, thereby forming aninterpenetrating polymer network.

The PFPE can include functional groups, such as functional groupsdisclosed herein. In some embodiments, the PFPE can include photocurablefunctional groups and/or thermalcurable functional groups. In someembodiments, where the PFPE includes a photocurable functional group,the PFPE can be cured by exposure to UV light after it has been wettedonto the ePTFE. The resulting membrane of ePTFE and PFPE includes, forexample, desirable flexibility, chemically resistance, and gaspermeability to solvent vapors and to air. The ePTFE portion of themembrane provides a continuous structure which serves to greatly toughenmaterials compared to just an elastomer itself. Some membranes aregenerally described by Zumbrum et al. in U.S. Pat. No. 6,673,455, whichis incorporated by reference herein in its entirety including allreference cited therein.

III. Method for Forming a Microfluidic Device Through a Thermal FreeRadical Curing Process

In some embodiments, the presently disclosed subject matter provides amethod for forming a microfluidic device by which a functional liquidperfluoropolyether (PFPE) precursor material is contacted with apatterned substrate, i.e., a master, and is thermally cured using a freeradical initiator. As provided in more detail herein below, in someembodiments, the liquid PFPE precursor material is fully cured to form afully cured PFPE network, which can then be removed from the patternedsubstrate and contacted with a second substrate to form a reversible,hermetic seal.

In some embodiments, the liquid PFPE precursor material is partiallycured to form a partially cured PFPE network. In some embodiments, thepartially cured network is contacted with a second partially cured layerof PFPE material and the curing reaction is taken to completion, therebyforming a permanent bond between the PFPE layers.

Further, the partially cured PFPE network can be contacted with a layeror substrate including another polymeric material, such aspoly(dimethylsiloxane) or another polymer, and then thermally cured sothat the PFPE network adheres to the other polymeric material.Additionally, the partially cured PFPE network can be contacted with asolid substrate, such as glass, quartz, or silicon, and then bonded tothe substrate through the use of a silane coupling agent.

III.A. Method of Forming a Patterned Layer of an Elastomeric Material

In some embodiments, the presently disclosed subject matter provides amethod of forming a patterned layer of an elastomeric material. Thepresently disclosed method is suitable for use with theperfluoropolyether material described in Sections II.A. and II.B., andthe fluoroolefin-based materials described in Section II.C. An advantageof using a higher viscosity PFPE material allows, among other things,for a higher molecular weight between cross links. A higher molecularweight between cross links can improve the elastomeric properties of thematerial, which can prevent among other things, cracks from forming.Referring now to FIGS. 1A-1C, a schematic representation of anembodiment of the presently disclosed subject matter is shown. Asubstrate 100 having a patterned surface 102 with a raised protrusion104 is depicted. Accordingly, the patterned surface 102 of the substrate100 includes at least one raised protrusion 104, which forms the shapeof a pattern. In some embodiments, patterned surface 102 of substrate100 includes a plurality of raised protrusions 104 which form a complexpattern.

As best seen in FIG. 1B, a liquid precursor material 106 is disposed onpatterned surface 102 of substrate 100. As shown in FIG. 1B, the liquidprecursor material 102 is treated with a treating process T_(r). Uponthe treating of liquid precursor material 106, a patterned layer 108 ofan elastomeric material (as shown in FIG. 1C) is formed.

As shown in FIG. 1C, the patterned layer 108 of the elastomeric materialincludes a recess 110 that is formed in the bottom surface of thepatterned layer 108. The dimensions of recess 110 correspond to thedimensions of the raised protrusion 104 of patterned surface 102 ofsubstrate 100. In some embodiments, recess 110 includes at least onechannel 112, which in some embodiments of the presently disclosedsubject matter includes a microscale channel. Patterned layer 108 isremoved from patterned surface 102 of substrate 100 to yieldmicrofluidic device 114. In some embodiments, removal of microfluidicdevice 114 is performed using a “lift-off” solvent which slowly wetsunderneath the device and releases it from the patterned substrate.Examples of such solvents include, but are not limited to, any solventthat will not adversely interact with the microfluidic device orfunctional components of the microfluidic device. Examples of suchsolvents include, but are not limited to: water, isopropyl alcohol,acetone, N-methylpyrollidinone, and dimethyl formamide.

In some embodiments, the patterned substrate includes an etched siliconwafer. In some embodiments, the patterned substrate includes aphotoresist patterned substrate. In some embodiments, the patternedsubstrate is treated with a coating that can aid in the release of thedevice from the patterned substrate or prevent reaction with latentgroups on a photoresist which constitutes the patterned substrate. Anexample of the coating can include, but is not limited to, a silane orthin film of metal deposited from a plasma, such as, a Gold/Palladiumcoating. For the purposes of the presently disclosed subject matter, thepatterned substrate can be fabricated by any of the processing methodsknown in the art, including, but not limited to, photolithography,electron beam lithography, and ion milling.

In some embodiments, the patterned layer of perfluoropolyether isbetween about 0.1 micrometers and about 100 micrometers thick. In someembodiments, the patterned layer of perfluoropolyether is between about0.1 millimeters and about 10 millimeters thick. In some embodiments, thepatterned layer of perfluoropolyether is between about one micrometerand about 50 micrometers thick. In some embodiments, the patterned layerof perfluoropolyether is about 20 micrometers thick. In someembodiments, the patterned layer of perfluoropolyether is about 5millimeters thick.

In some embodiments, the patterned layer of perfluoropolyether includesa plurality of microscale channels. In some embodiments, the channelshave a width ranging from about 0.01 μm to about 1000 μm; a widthranging from about 0.05 μm to about 1000 μm; and/or a width ranging fromabout 1 μm to about 1000 μm. In some embodiments, the channels have awidth ranging from about 1 μm to about 500 μm; a width ranging fromabout 1 μm to about 250 μm; and/or a width ranging from about 10 μm toabout 200 μm. Exemplary channel widths include, but are not limited to,0.1 μm, 1 μm, 2 μm, 5 μm, 10 μm, 20 μm, 30 μm, 40 μm, 50 μm, 60 μm, 70μm, 80 μm, 90 μm, 100 μm, 110 μm, 120 μm, 130 μm, 140 μm, 150 μm, 160μm, 170 μm, 180 μm, 190 μm, 200 μm, 210 μm, 220 μm, 230 μm, 240 μm, and250 μm.

In some embodiments, the channels have a depth ranging from about 1 μmto about 1000 μm; and/or a depth ranging from about 1 μm to 100 μm. Insome embodiments, the channels have a depth ranging from about 0.01 μmto about 1000 μm; a depth ranging from about 0.05 μm to about 500 μm; adepth ranging from about 0.2 μm to about 250 μm; a depth ranging fromabout 1 μm to about 100 μm; a depth ranging from about 2 μm to about 20μm; and/or a depth ranging from about 5 μm to about 10 μm. Exemplarychannel depths include, but are not limited to, 0.01 μm, 0.02 μm, 0.05μm, 0.1 μm, 0.2 μm, 0.5 μm, 1 μm, 2 μm, 3 μm, 4 μm, 5 μm, 7.5 μm, 10 μm,12.5 μm, 15 μm, 17.5 μm, 20 μm, 22.5 μm, 25 μm, 30 μm, 40 μm, 50 μm, 75μm, 100 μm, 150 μm, 200 μm, and 250 μm.

In some embodiments, the channels have a width-to-depth ratio rangingfrom about 0.1:1 to about 100:1. In some embodiments, the channels havea width-to-depth ratio ranging from about 1:1 to about 50:1. In someembodiments, the channels have a width-to-depth ratio ranging from about2:1 to about 20:1. In some embodiments, the channels have awidth-to-depth ratio ranging from about 3:1 to about 15:1. In someembodiments, the channels have a width-to-depth ratio of about 10:1.

One of ordinary skill in the art would recognize that the dimensions ofthe channels of the presently disclosed subject matter are not limitedto the exemplary ranges described hereinabove and can vary in width anddepth to affect the magnitude of force required to flow a materialthrough the channel and/or to actuate a valve to control the flow of thematerial therein. Further, as will be described in more detail hereinbelow, channels of greater width are contemplated for use as a fluidreservoir, a reaction chamber, a mixing channel, a separation region,and the like.

III.B. Method for Forming a Multilayer Patterned Material

In some embodiments, the presently disclosed subject matter describes amethod for forming a multilayer patterned material, e.g., a multilayerpatterned PFPE material. In some embodiments, the multilayer patternedperfluoropolyether material is used to fabricate a monolithic PFPE-basedmicrofluidic device.

Referring now to FIGS. 2A-2D, a schematic representation of thepreparation of an embodiment of the presently disclosed subject matteris shown. Patterned layers 200 and 202 are provided, each of which, insome embodiments, include a perfluoropolyether material prepared from aliquid PFPE precursor material having a viscosity greater than about 100cSt. In this example, each of the patterned layers 200 and 202 include aplurality of channels 204. In this embodiment of the presently disclosedsubject matter, the plurality of channels 204 include microscalechannels. In patterned layer 200, the channels are represented by adashed line, i.e., in shadow, in FIGS. 2A-2C. Patterned layer 202 isoverlaid on patterned layer 200 in a predetermined alignment. In thisexample, the predetermined alignment is such that channels 204 inpatterned layer 200 and 202 are substantially perpendicular to eachother. In some embodiments, as depicted in FIGS. 2A-2D, patterned layer200 is overlaid on nonpatterned layer 206. Nonpatterned layer 206 caninclude a perfluoropolyether.

Continuing with reference to FIGS. 2A-2D, patterned layers 200 and 202,and in some embodiments nonpatterned layer 206, are treated by atreating process T_(r). As described in more detail herein below, layers200, 202, and, in some embodiments nonpatterned layer 206, are treatedby treating T_(r), to promote the adhesion of patterned layers 200 and202 to each other, and in some embodiments, patterned layer 200 tononpatterned layer 206, as shown in FIGS. 2C and 2D. The resultingmicrofluidic device 208 includes an integrated network 210 of microscalechannels 204 which intersect predetermined intersecting points 212, asbest seen in the cross-section provided in FIG. 2D. Also shown in FIG.2D is membrane 214 comprising the top surface of channels 204 ofpatterned layer 200 which separates channel 204 of patterned layer 202from channels 204 of patterned layer 200.

Continuing with reference to FIGS. 2A-2C, in some embodiments, patternedlayer 202 includes a plurality of apertures, and the apertures aredesignated input aperture 216 and output aperture 218. In someembodiments, the holes, e.g., input aperture 216 and output aperture 218are in fluid communication with channels 204. In some embodiments, theapertures include a side-actuated valve structure constructed of, forexample, a thin membrane of PFPE material which can be actuated torestrict the flow in an abutting channel. It will be appreciated,however, that the side-actuated valve structure can be constructed fromother materials disclosed herein.

In some embodiments, the first patterned layer of photocured PFPEmaterial is cast at such a thickness to impart a degree of mechanicalstability to the PFPE structure. Accordingly, in some embodiments, thefirst patterned layer of the photocured PFPE material is about 50 μm toseveral centimeters thick. In some embodiments, the first patternedlayer of the photocured PFPE material is between 50 μm and about 10millimeters thick. In some embodiments, the first patterned layer of thephotocured PFPE material is 5 mm thick. In some embodiments, the firstpatterned layer of PFPE material is about 4 mm thick. Further, in someembodiments, the thickness of the first patterned layer of PFPE materialranges from about 0.1 μm to about 10 cm; from about 1 μm to about 5 cm;from about 10 μm to about 2 cm; and from about 100 μm to about 10 mm.

In some embodiments, the second patterned layer of the photocured PFPEmaterial is between about 1 micrometer and about 100 micrometers thick.In some embodiments, the second patterned layer of the photocured PFPEmaterial is between about 1 micrometer and about 50 micrometers thick.In some embodiments, the second patterned layer of the photocuredmaterial is about 20 micrometers thick.

Although FIGS. 2A-2C disclose the formation of a microfluidic devicewherein two patterned layers of PFPE material are combined, in someembodiments of the presently disclosed subject matter it is possible toform a microfluidic device having one patterned layer and onenon-patterned layer of PFPE material. Thus, the first patterned layercan include a microscale channel or an integrated network of microscalechannels and then the first patterned layer can be overlaid on top ofthe non-patterned layer and adhered to the non-patterned layer using aphotocuring step, such as application of ultraviolet light as disclosedherein, to form a monolithic structure including enclosed channelstherein.

Accordingly, in some embodiments, a first and a second patterned layerof photocured perfluoropolyether material, or alternatively a firstpatterned layer of photocured perfluoropolyether material and anonpatterned layer of photocured perfluoropolyether material, adhere toone another, thereby forming a monolithic PFPE-based microfluidicdevice.

III.C. Method of Forming a Patterned PFPE Layer Through a Thermal FreeRadical Curing Process

In some embodiments, a thermal free radical initiator, including, butnot limited to, a peroxide and/or an azo compound, is blended with aliquid perfluoropolyether (PFPE) precursor, which is functionalized witha polymerizable group, including, but not limited to, an acrylate, amethacrylate, and a styrenic unit to form a blend. As shown in FIGS.1A-1C, the blend is then contacted with a patterned substrate, i.e., a“master,” and heated to cure the PFPE precursor into a network.

In some embodiments, the PFPE precursor is fully cured to form a fullycured PFPE precursor. In some embodiments, the free radical curingreaction is allowed to proceed only partially to form a partially-curednetwork.

III.D. Method of Adhering a Layer of a PFPE Material to a SubstrateThrough a Thermal Free Radical Curing Process

In some embodiments the fully cured PFPE precursor is removed, e.g.,peeled, from the patterned substrate, i.e., the master, and contactedwith a second substrate to form a reversible, hermetic seal.

In some embodiments, the partially cured network is contacted with asecond partially cured layer of PFPE material and the curing reaction istaken to completion, thereby forming a permanent bond between the PFPElayers.

In some embodiments, the partial free-radical curing method is used tobond at least one layer of a partially-cured PFPE material to asubstrate. In some embodiments, the partial free-radical curing methodis used to bond a plurality of layers of a partially-cured PFPE materialto a substrate. In some embodiments, the substrate is selected from thegroup consisting of a glass material, a quartz material, a siliconmaterial, a fused silica material, and a plastic material. In someembodiments, the substrate is treated with a silane coupling agent.

An embodiment of the presently disclosed method for adhering a layer ofPFPE material to a substrate is illustrated in FIGS. 3A-3C. Referringnow to FIG. 3A, a substrate 300 is provided, wherein, in someembodiments, substrate 300 is selected from the group consisting of aglass material, a quartz material, a silicon material, a fused silicamaterial, and a plastic material. Substrate 300 is treated by treatingprocess T_(r1). In some embodiments, treating process T_(r1) includestreating the substrate with a base/alcohol mixture, e.g.,KOH/isopropanol, to impart a hydroxyl functionality to substrate 300.

Referring now to FIG. 3B, functionalized substrate 300 is reacted with asilane coupling agent, e.g., R—SiCl₃ or R—Si(OR₁)₃, wherein R and R₁represent a functional group as described herein to form a silanizedsubstrate 300. In some embodiments, the silane coupling agent isselected from the group consisting of a monohalosilane, a dihalosilane,a trihalosilane, a monoalkoxysilane, a dialkoxysilane, and atrialkoxysilane; and wherein the monohalosilane, dihalosilane,trihalosilane, monoalkoxysilane, dialkoxysilane, and trialkoxysilane arefunctionalized with a moieties selected from the group consisting of anamine, a methacrylate, an acrylate, a styrenic, an epoxy, an isocyanate,a halogen, an alcohol, a benzophenone derivative, a maleimide, acarboxylic acid, an ester, an acid chloride, and an olefin.

Referring now to FIG. 3C, silanized substrate 300 is contacted with apatterned layer of partially cured PFPE material 302 and treated bytreating process T_(r2) to form a permanent bond between patterned layerof PFPE material 302 and substrate 300.

In some embodiments, a partial free radical cure is used to adhere aPFPE layer to a second polymeric material, such as a poly(dimethylsiloxane) (PDMS) material, a polyurethane material, asilicone-containing polyurethane material, and a PFPE-PDMS blockcopolymer material. In some embodiments, the second polymeric materialincludes a functionalized polymeric material. In some embodiments, thesecond polymeric material is encapped with a polymerizable group. Insome embodiments, the polymerizable group is selected from the groupconsisting of an acrylate, a styrene, and a methacrylate. Further, insome embodiments, the second polymeric material is treated with a plasmaand a silane coupling agent to introduce the desired functionality tothe second polymeric material.

An embodiment of the presently disclosed method for adhering a patternedlayer of PFPE material to another patterned layer of polymeric materialis illustrated in FIGS. 4A-4C. Referring now to FIG. 4A, a patternedlayer of a first polymeric material 400 is provided. In someembodiments, first polymeric material includes a PFPE material. In someembodiments, first polymeric material includes a polymeric materialselected from the group consisting of a poly(dimethylsiloxane) material,a polyurethane material, a silicone-containing polyurethane material,and a PFPE-PDMS block copolymer material. Patterned layer of firstpolymeric material 400 is treated by treating process T_(r1). In someembodiments, treating process T_(r1) includes exposing the patternedlayer of first polymeric material 400 to UV light in the presence of O₃and an R functional group, to add an R functional group to the patternedlayer of polymeric material 400.

Referring now to FIG. 4B, the functionalized patterned layer of firstpolymeric material 400 is contacted with the top surface of afunctionalized patterned layer of PFPE material 402 and then treated bytreating process T_(r2) to form a two layer hybrid assembly 404. Thus,functionalized patterned layer of first polymeric material 400 isthereby bonded to functionalized patterned layer of PFPE material 402.

Referring now to FIG. 4C, two-layer hybrid assembly 404, in someembodiments, is contacted with substrate 406 to form a multilayer hybridstructure 410. In some embodiments, substrate 406 is coated with a layerof liquid PFPE precursor material 408. Multilayer hybrid structure 410is treated by treating process T_(r3) to bond two-layer assembly 404 tosubstrate 406.

IV. Methods for Forming a Device Through a Two-Component Curing Process

The presently disclosed subject matter provides a method for forming adevice by which a polymer, such as, functional perfluoropolyether (PFPE)precursors, are contacted with a patterned surface and then curedthrough the reaction of two components, such as epoxy/amine,epoxy/hydroxyl, triol/diisocyanate, carboxylic acid/amine, carboxylicacid/hydroxyl, ester/amine, ester/hydroxyl, amine/anhydride, acidhalide/hydroxyl, acid halide/amine, amine/halide, hydroxyl/halide,hydroxyl/chlorosilane, azide/acetylene and other so-called “clickchemistry” reactions, and metathesis reactions involving the use ofGrubb's-type catalysts to form a fully-cured or a partially-cured PFPEnetwork. Examples of UV and thermal end-cap combinations, according tosome embodiments, include: UV diurethane methacrylate with thermal trioland diisocyanate components, UV diurethane methacrylate with thermaltetrol and diisocyanate components, UV diurethane methacrylate with UVdiepoxy, UV diurethane methacrylate with thermal diepoxy and diaminecomponents, UV diurethane methacrylate with thermal diisocyanate,combinations thereof, and the like.

As used herein the term “click chemistry” refers to a term used in theart to describe the synthesis of compounds using any of a number ofcarbon-heteroatom bond forming reactions. “Click chemistry” reactionstypically are relatively insensitive to oxygen and water, have highstereoselectivity and yield, and thermodynamic driving forces of about20 kcal/mol or greater. In some embodiments, useful “click chemistry”reactions include cycloaddition reactions of unsaturated compounds,including 1,3-dipolar additions and Diels-Alder reactions; nucleophilicsubstitution reactions, especially those involving ring opening ofsmall, strained rings like epoxides and aziridines; addition reactionsto carbon-carbon multiple bonds; and reactions involving non-aldolcarbonyl chemistry, such as the formation of ureas and amides.

Further, the term “metathesis reactions” refers to reactions in whichtwo compounds react to form two new compounds with no change inoxidation numbers in the final products. For example, olefin metathesisinvolves the 2+2 cycloaddition of an olefin and a transition metalalkylidene complex to form a new olefin and a new alkylidene. Inring-opening metathesis polymerization (ROMP), the olefin is a strainedcyclic olefin, and 2+2 cycloaddition to the transition metal catalystinvolves opening of the strained ring. The growing polymer remains partof the transition metal complex until capped, for example, by 2+2cycloaddition to an aldehyde. Grubbs catalysts for metathesis reactionswere first described in 1996 (see Schwab, P., et al., J. Am. Chem. Soc.,118, 100-110 (1996)). Grubbs catalysts are transition metal alkylidenescontaining ruthenium supported by phosphine ligands and are unique inthat that they are tolerant of different functionalities in the alkeneligand.

Accordingly, in one embodiment, the photocurable component can includefunctional groups that can undergo photochemical 2+2 cycloadditions.Such groups include alkenes, aldehydes, ketones, and alkynes.Photochemical 2+2 cycloadditions can be used, for example, to formcyclobutanes and oxetanes.

Thus, in some embodiments, the partially-cured PFPE network is contactedwith another substrate, and the curing is then taken to completion toadhere the PFPE network to the substrate. This method can be used toadhere multiple layers of a PFPE material to a substrate.

Further, in some embodiments, the substrate includes a second polymericmaterial, such as PDMS, or another polymer. In some embodiments, thesecond polymeric material includes an elastomer other than PDMS, such asKratons™ (Shell Chemical Company), buna rubber, natural rubber, afluoroelastomer, chloroprene, butyl rubber, nitrile rubber,polyurethane, or a thermoplastic elastomer. In some embodiments, thesecond polymeric material includes a rigid thermoplastic material,including but not limited to: polystyrene, poly(methyl methacrylate), apolyester, such as poly(ethylene terephthalate), a polycarbonate, apolyimide, a polyamide, a polyvinylchloride, a polyolefin, apoly(ketone), a poly(ether ether ketone), and a poly(ether sulfone).

In some embodiments, the PFPE layer is adhered to a solid substrate,such as a glass material, a quartz material, a silicon material, and afused silica material, through use of a silane coupling agent.

IV.A. Method of Forming a Patterned PFPE Layer Through a Two-ComponentCuring Process

In some embodiments, a PFPE network is formed through the reaction of atwo-component functional liquid precursor system. Using the generalmethod for forming a patterned layer of polymeric material as shown inFIGS. 1A-1C, a liquid precursor material that includes a two-componentsystem is contacted with a patterned substrate and a patterned layer ofPFPE material is formed. In some embodiments, the two-component liquidprecursor system is selected from the group consisting of anepoxy/amine, epoxy/hydroxyl, triol/diisocyanate, carboxylic acid/amine,carboxylic acid/hydroxyl, ester/amine, ester/hydroxyl, amine/anhydride,acid halide/hydroxyl, acid halide/amine, amine/halide, hydroxyl/halide,hydroxyl/chlorosilane, azide/acetylene and other so-called “clickchemistry” reactions, and metathesis reactions involving the use ofGrubb's-type catalysts. The functional liquid precursors are blended inthe appropriate ratios and then contacted with a patterned surface ormaster. The curing reaction is allowed to take place by using heat,catalysts, and the like, until the network is formed.

In some embodiments, a fully cured PFPE precursor is formed. In someembodiments, the two-component reaction is allowed to proceed onlypartially, thereby forming a partially cured PFPE network.

IV.B. Method of Adhering a PFPE Layer to a Substrate Through aTwo-Component Curing Process

IV.B.1. Full Cure with a Two-Component Curing Process

In some embodiments, the fully cured PFPE two-component precursor isremoved, e.g., peeled, from the master and contacted with a substrate toform a reversible, hermetic seal. In some embodiments, the partiallycured network is contacted with another partially cured layer of PFPEand the reaction is taken to completion, thereby forming a permanentbond between the layers. In some embodiments, the cure component can bea photo-cure component that is activated upon exposure to photo or UVradiation. In alternative embodiments, the cure component can be athermal-cure component that is activated upon the application of thermalenergy. In yet alternative embodiments, the cure component can beactivated by a moisture cure mechanism. Moisture cure mechanisms aredisclosed in S. Turri, et al. Surf. Interface Anal. 29, 873-886, whichis incorporated herein by reference in its entirety including allreferences cited therein.

IV.B.2. Partial Cure with a Two-Component System

As shown in FIGS. 3A-3C, in some embodiments, the partial two-componentcuring method is used to bond at least one layer of a partially-curedPFPE material to a substrate. In some embodiments, the partialtwo-component curing method is used to bond a plurality of layers of apartially-cured PFPE material to a substrate. In some embodiments, thesubstrate is selected from the group consisting of a glass material, aquartz material, a silicon material, a fused silica material, and aplastic material. In some embodiments, the substrate is treated with asilane coupling agent.

As shown in FIGS. 4A-4C, in some embodiments, a partial two-componentcure is used to adhere the PFPE layer to a second polymeric material,such as a poly(dimethylsiloxane) (PDMS) material. In some embodiments,the PDMS material includes a functionalized PDMS material. In someembodiments, the PDMS is treated with a plasma and a silane couplingagent to introduce the desired functionality to the PDMS material. Insome embodiments, the PDMS material is encapped with a polymerizablegroup. In some embodiments, the polymerizable group includes an epoxide.In some embodiments, the polymerizable group includes an amine.

In some embodiments, the second polymeric material includes an elastomerother than PDMS, such as Kratons™, buna rubber, natural rubber, afluoroelastomer, chloroprene, butyl rubber, nitrile rubber,polyurethane, or a thermoplastic elastomer. In some embodiments, thesecond polymeric material includes a rigid thermoplastic, including butnot limited to: polystyrene, poly(methyl methacrylate), a polyester,such as poly(ethylene terephthalate), a polycarbonate, a polyimide, apolyamide, a polyvinylchloride, a polyolefin, a poly(ketone), apoly(ether ether ketone), and a poly(ether sulfone).

IV.B.3. Excess Cure with a Two-Component System

The presently disclosed subject matter provides a method for forming amicrofluidic device by which a functional perfluoropolyether (PFPE)precursor is contacted with a patterned substrate and cured through thereaction of two components, such as epoxy/amine, epoxy/hydroxyl,triol/diisocyanate, carboxylic acid/amine, carboxylic acid/hydroxyl,ester/amine, ester/hydroxyl, amine/anhydride, acid halide/hydroxyl, acidhalide/amine, amine/halide, hydroxyl/halide, hydroxyl/chlorosilane,azide/acetylene and other so-called “click chemistry” reactions, andmetathesis reactions involving the use of Grubb's-type catalysts, toform a layer of cured PFPE material. In this particular method, thelayer of cured PFPE material can be adhered to a second substrate byfully curing the layer with an excess of one component and contactingthe layer of cured PFPE material with a second substrate having anexcess of a second component in such a way that the excess groups reactto adhere the layers.

Thus, in some embodiments, a two-component system, such as anepoxy/amine, epoxy/hydroxyl, triol/diisocyanate, carboxylic acid/amine,carboxylic acid/hydroxyl, ester/amine, ester/hydroxyl, amine/anhydride,acid halide/hydroxyl, acid halide/amine, amine/halide, hydroxyl/halide,hydroxyl/chlorosilane, azide/acetylene and other so-called “clickchemistry” reactions, and metathesis reactions involving the use ofGrubb's-type catalysts, is blended. In some embodiments, at least onecomponent of the two-component system is in excess of the othercomponent. The reaction is then taken to completion by heating, using acatalyst, and the like, with the remaining cured network having aplurality of functional groups generated by the presence of the excesscomponent.

In some embodiments, two layers of fully cured PFPE materials includingcomplimentary excess groups are contacted with one another, wherein theexcess groups are allowed to react, thereby forming a permanent bondbetween the layers.

As shown in FIGS. 3A-3C, in some embodiments, a fully cured PFPE networkincluding excess functional groups is contacted with a substrate. Insome embodiments, the substrate is selected from the group consisting ofa glass material, a quartz material, a silicon material, a fused silicamaterial, and a plastic material. In some embodiments, the substrate istreated with a silane coupling agent such that the functionality on thecoupling agent is complimentary to the excess functionality on the fullycured network. Thus, a permanent bond is formed to the substrate.

As shown in FIGS. 4A-4C, in some embodiments, the two-component excesscure is used to bond a PFPE network to a second polymeric material, suchas a poly(dimethylsiloxane) PDMS material. In some embodiments, the PDMSmaterial includes a functionalized PDMS material. In some embodiments,the PDMS material is treated with a plasma and a silane coupling agentto introduce the desired functionality. In some embodiments, the PDMSmaterial is encapped with a polymerizable group. In some embodiments,the polymerizable material includes an epoxide. In some embodiments, thepolymerizable material includes an amine.

In some embodiments, the second polymeric material includes an elastomerother than PDMS, such as Kratons™, buna rubber, natural rubber,a-fluoroelastomer, chloroprene, butyl rubber, nitrile rubber,polyurethane, or a thermoplastic elastomer. In some embodiments, thesecond polymeric material includes a rigid thermoplastic, including butnot limited to: polystyrene, poly(methyl methacrylate), a polyester,such as poly(ethylene terephthalate), a polycarbonate, a polyimide, apolyamide, a polyvinylchloride, a polyolefin, a poly(ketone), apoly(ether ether ketone), and a poly(ether sulfone).

IV.B.4. Blending a Thermalcurable Component with a Photocurable Material

According to yet another embodiment, microfluidic devices are formedfrom adhering multiple layers of materials together. In one embodiment,a two-component thermally curable material is blended with aphotocurable material, thereby creating a multiple stage curingmaterial. In certain embodiments, the two-component system can includefunctional groups, such as epoxy/amine, epoxy/hydroxyl,triol/diisocyanate, carboxylic acid/amine, carboxylic acid/hydroxyl,ester/amine, ester/hydroxyl, amine/anhydride, acid halide/hydroxyl, acidhalide/amine, amine/halide, hydroxyl/halide, hydroxyl/chlorosilane,azide/acetylene and other so-called “click chemistry” reactions, andmetathesis reactions involving the use of Grubb's-type catalysts. In oneembodiment, the photocurable component can include such functionalgroups as: acrylates, styrenics, epoxides, cyclobutanes and other 2+2cycloadditions.

In some embodiments, a two-component thermally curable material isblended in varying ratios with a photocurable material. In oneembodiment, the material can then be deposited on a patterned substrateas described above. Such a system can be exposed to actinic radiation,e.g., UV light, and solidified into a network, while the thermallycurable components are mechanically entangled in the network but remainunreacted. Layers of the material can then be prepared, for example,cut, trimmed, punched with inlet/outlet holes, and aligned inpredetermined positions on a second, photocured layer. Once thephotocured layers are aligned and sealed, the device can be heated toactivate the thermally curable component within the layers. When thethermally curable components are activated by the heat, the layers areadhered together by reaction at the interface.

In some embodiments, the thermal reaction is taken to completion. Inother embodiments, the thermal reaction is initially only done partiallyand after multiple layers are aligned, the thermal reaction is taken tocompletion thereby adhering the layers together. In other embodiments, amultilayered device is formed and adhered to a final flat, non-patternedlayer through the thermal cure.

In some embodiments, the thermal cure reaction is done first. The layeris then prepared, for example, cut, trimmed, punched with inlet/outletholes, and aligned. Next, the photocurable component is activated byexposure to actinic radiation, e.g., UV light, and the layers areadhered by functional groups reacting at the interface between thelayers.

In some embodiments, blended two-component thermally curable andphotocurable materials are used to bond a PFPE network to a secondpolymeric material, such as a poly(dimethylsiloxane) PDMS material. Insome embodiments, the PDMS material includes a functionalized PDMSmaterial. As will be appreciated by one of ordinary skill in the art,the functionalized PDMS material is PDMS material that contains areactive chemical group, as described elsewhere herein. In someembodiments, the PDMS material is treated with a plasma and a silanecoupling agent to introduce the desired functionality. In someembodiments, the PDMS material is encapped with a polymerizable group.In some embodiments, the polymerizable material includes an epoxide. Insome embodiments, the polymerizable material includes an amine.

In some embodiments, the second polymeric material includes an elastomerother than PDMS, such as Kratons™, buna rubber, natural rubber, afluoroelastomer, chloroprene, butyl rubber, nitrile rubber,polyurethane, or a thermoplastic elastomer. In some embodiments, thesecond polymeric material includes a rigid thermoplastic, including butnot limited to: polystyrene, poly(methyl methacrylate), a polyester,such as poly(ethylene terephthalate), a polycarbonate, a polyimide, apolyamide, a polyvinylchloride, a polyolefin, a poly(ketone), apoly(ether ether ketone), and a poly(ether sulfone).

In some embodiments, a blend of a photocurable PFPE liquid precursor anda two-component thermally curable PFPE liquid precursor is made in sucha way that one component of the two component thermally curable blend isin excess of the other. In this way, multiple layers can be adheredthrough residual complimentary functional groups present in multiplelayers.

According to a preferred embodiment, the amount of thermal cure andphotocure substance added to the material is selected to produceadhesion between layers of the completed device that can withstand apressure up to a desired pressure per square inch. According to someembodiments, the amount of thermal cure and photocure substance added tothe material is selected to produce adhesion between layers of thedevice that can withstand pressures up to about 120 psi. According toother embodiments, the laminated device can withstand pressure up toabout 110 psi. According to other embodiments, the laminated device canwithstand pressure up to about 100 psi. According to other embodiments,the laminated device can withstand pressure up to about 90 psi.According to other embodiments, the laminated device can withstandpressure up to about 80 psi. According to other embodiments, thelaminated device can withstand pressure up to about 70 psi. Inalternative embodiment the adhesion between layers is structured suchthat a device can withstand pressures between about 5 psi and about 120psi in different ranges of the device without delamination. According toyet a further embodiment, the amount of thermal cure and photocuresubstance added to the material is selected to produce adhesion betweenlayers of the device that can withstand pressures between about 10 psiand about 60 psi without delaminating. An example of a valve in a PFPEdevice actuated at about 45 psi is shown in FIG. 12. No delamination isobserved after cycling the valve repeatedly for extended periods oftime.

An illustrative example of a method for making a multilayeredmicrofluidic device will now be described with respect to FIGS. 11 a-11e. A two-component thermally curable material blended with aphotocurable material is disposed on patterned templates 5006, 5008(sometimes referred to as a master template or template), as shown inFIG. 11 a. According to alternative embodiments of the presentinvention, the blended material can be spin coated onto the patternedtemplate or cast onto the patterned template by pooling the materialinside a gasket. In some embodiments, spin coating is used to form thinlayers such as first layer 5002 and a cast technique is used to formthick layers such as second layer 5004, as will be appreciated by one ofordinary skill in the art. Next, the blended material positioned ontemplates 5006 and 5008 is treated with an initial cure, such as aphotocure, to form first layer 5002 and second layer 5004, respectively.The photocure partially cures the material but does not initiate thethermal cure components of the material. Patterned template 5008 is thenremoved from second layer 5004. Removal of patterned templates from thelayers is described in more detail herein. Next, second layer 5004 ispositioned with respect to first layer 5002 and the combination istreated with a second cure, as shown in FIG. 11 b, which results in thebonding, or adhesion, between first layer 5002 and second layer 5004,collectively referred to hereinafter as the “two adhered layers 5002 and5004.” In some embodiments, the second cure is an initial heat curingthat initiates the two-component thermal cure of the material. Next, thetwo adhered layers 5002 and 5004 are removed from patterned template5006, as shown in FIG. 11 c. In FIG. 11 d, the two adhered layers 5002and 5004 are positioned on flat layer 5014, flat layer 5014 previouslybeing coated onto flat template 5012 and treated with an initial cure.The combination of layers 5002, 5004, and 5014 is then treated to afinal cure to fully adhere all three layers together, as shown in FIG.11 e.

According to alternative embodiments, patterned template 5006 can becoated with release layer 5010 to facilitate removal of the cured orpartially cured layers (see FIG. 11 c). Further, coating of thetemplates, e.g., patterned template 5006 and/or patterned template 5008,can reduce reaction of the thermal components with latent groups presenton the template. For example, release layer 5010 can be a Gold/Palladiumcoating.

According to alternative embodiments, removal of the partially cured andcured layers can be realized by peeling, suction, pneumatic pressure,through the application of solvents to the partially cured or curedlayers, or through a combination of these teachings.

V. Method for Functionalizing a Surface of a Micro- and/or Nano-scaleDevice

In some embodiments, the presently disclosed subject matter providesmaterials and methods for functionalizing the channels in a microfluidicdevice and/or a microtiter well. In some embodiments, suchfunctionalization includes, but is not limited to, the synthesis and/orattachment of peptides and other natural polymers to the interiorsurface of a channel in a microfluidic device. Accordingly, thepresently disclosed subject matter can be applied to microfluidicdevices, such as those described by Rolland, J., et al., JACS 2004, 126,2322-2323, the disclosure of which is incorporated herein by referencein its entirety.

In some embodiments, the method includes binding a small molecule to theinterior surface of a microfluidic channel or the surface of amicrotiter well. In such embodiments, once bound, the small molecule canserve a variety of functions. In some embodiments, the small moleculefunctions as a cleavable group, which when activated, can change thepolarity of the channel and hence the wettability of the channel. Insome embodiments, the small molecule functions as a binding site. Insome embodiments, the small molecule functions as a binding site for oneof a catalyst, a drug, a substrate for a drug, an analyte, and a sensor.In some embodiments, the small molecule functions as a reactivefunctional group. In some embodiments, the reactive functional group isreacted to yield a zwitterion. In some embodiments, the zwitterionprovides a polar, ionic channel.

An embodiment of the presently disclosed method for functionalizing theinterior surface of a microfluidic channel and/or a microtiter well isillustrated in FIGS. 5A and 5B. Referring now to FIG. 5A, a microfluidicchannel 500 is provided. In some embodiments, microfluidic channel 500is formed from a functional PFPE material having an R functional group,as described herein. In some embodiments, microchannel 500 includes aPFPE network which undergoes a post-curing treating process, wherebyfunctional group R is introduced into the interior surface 502 ofmicrofluidic channel 500.

Referring now to FIG. 5B, a microtiter well 504 is provided. In someembodiments, microtiter well 504 is coated with a layer offunctionalized PFPE material 506, which includes an R functional group,to impart functionality into microtiter well 504.

V.A. Method of Attaching a Functional Group to a PFPE Network

In some embodiments, PFPE networks including excess functionality areused to functionalize the interior surface of a microfluidic channel orthe surface of a microtiter well. In some embodiments, the interiorsurface of a microfluidic channel or the surface of a microtiter well isfunctionalized by attaching a functional moiety selected from the groupconsisting of a protein, an oligonucleotide, a drug, a ligand, acatalyst, a dye, a sensor, an analyte, and a charged species capable ofchanging the wettability of the channel.

In some embodiments, latent functionalities are introduced into thefully cured PFPE network. In some embodiments, latent methacrylategroups are present at the surface of the PFPE network that has been freeradically cured either photochemically or thermally. Multiple layers offully cured PFPE are then contacted with the functionalized surface ofthe PFPE network, forming a seal, and reacted, by heat, for example, toallow the latent functionalities to react and form a permanent bondbetween the layers.

In some embodiments, the latent functional groups react photochemicallywith one another at a wavelength different from that used to cure thePFPE precursors. In some embodiments, this method is used to adherefully cured layers to a substrate. In some embodiments, the substrate isselected from the group consisting of a glass material, a quartzmaterial, a silicon material, a fused silica material, and a plasticmaterial. In some embodiments, the substrate is treated with a silanecoupling agent complimentary to the latent functional groups.

In some embodiments, such latent functionalities are used to adhere afully cured PFPE network to a second polymeric material, such as apoly(dimethylsiloxane) PDMS material. In some embodiments, the PDMSmaterial includes a functionalized PDMS material. In some embodiments,the PDMS material is treated with a plasma and a silane coupling agentto introduce the desired functionality. In some embodiments, the PDMSmaterial is encapped with a polymerizable group. In some embodiments,the polymerizable group is selected from the group consisting of anacrylate, a styrene, and a methacrylate.

In some embodiments, the second polymeric material includes an elastomerother than PDMS, such as Kratons™, buna rubber, natural rubber, afluoroelastomer, chloroprene, butyl rubber, nitrile rubber,polyurethane, or a thermoplastic elastomer. In some embodiments, thesecond polymeric material includes a rigid thermoplastic, including butnot limited to: polystyrene, poly(methyl methacrylate), a polyester,such as poly(ethylene terephthalate), a polycarbonate, a polyimide, apolyamide, a polyvinylchloride, a polyolefin, a poly(ketone), apoly(ether ether ketone), and a poly(ether sulfone).

V.B. Method of Introducing Functionality in the Generation of a LiquidPFPE Precursor

The presently disclosed subject matter provides a method of forming amicrofluidic device by which a photochemically cured PFPE layer isplaced in conformal contact with a second substrate thereby forming aseal. The PFPE layer is then heated at elevated temperatures to adherethe layer to the substrate through latent functional groups. In someembodiments, the second substrate also includes a cured PFPE layer. Insome embodiments, the second substrate includes a second polymericmaterial, such as a poly(dimethylsiloxane) (PDMS) material.

In some embodiments, the second polymeric material includes an elastomerother than PDMS, such as Kratons™, buna rubber, natural rubber, afluoroelastomer, chloroprene, butyl rubber, nitrile rubber,polyurethane, or a thermoplastic elastomer. In some embodiments, thesecond polymeric material includes a rigid thermoplastic, including butnot limited to: polystyrene, poly(methyl methacrylate), a polyester,such as poly(ethylene terephthalate), a polycarbonate, a polyimide, apolyamide, a polyvinylchloride, a polyolefin, a poly(ketone), apoly(ether ether ketone), and a poly(ether sulfone).

In some embodiments, the latent groups include methacrylate units thatare not reacted during the photocuring process. Further, in someembodiments, the latent groups are introduced in the generation of theliquid PFPE precursor. For example, in some embodiments, methacrylateunits are added to a PFPE diol through the use of glycidyl methacrylate,the reaction of the hydroxy and the epoxy group generates a secondaryalcohol, which can be used as a handle to introduce chemicalfunctionality. In some embodiments, multiple layers of fully cured PFPEare adhered to one another through these latent functional groups. Insome embodiments, the latent functionalities are used to adhere a fullycured PFPE layer to a substrate. In some embodiments, the substrate isselected from the group consisting of a glass material, a quartzmaterial, a silicon material, a fused silica material, and a plasticmaterial. In some embodiments, the substrate is treated with a silanecoupling agent.

Further, this method can be used to adhere a fully cured PFPE layer to asecond polymeric material, such as a poly(dimethylsiloxane) (PDMS)material. In some embodiments, the PDMS material includes afunctionalized PDMS material. In some embodiments, the PDMS material istreated with a plasma and a silane coupling agent to introduce thedesired functionality. In some embodiments, the PDMS material isencapped with a polymerizable group. In some embodiments, thepolymerizable material is selected from the group consisting of anacrylate, a styrene, and a methacrylate.

In some embodiments, the second polymeric material includes an elastomerother than PDMS, such as Kratons™, buna rubber, natural rubber, afluoroelastomer, chloroprene, butyl rubber, nitrile rubber,polyurethane, or a thermoplastic elastomer. In some embodiments, thesecond polymeric material includes a rigid thermoplastic, including butnot limited to: polystyrene, poly(methyl methacrylate), a polyester,such as poly(ethylene terephthalate), a polycarbonate, a polyimide, apolyamide, a polyvinylchloride, a polyolefin, a poly(ketone), apoly(ether ether ketone), and a poly(ether sulfone).

In some embodiments, PFPE networks containing latent functionality areused to functionalize the interior surface of a microfluidic channel ora microtiter well. Examples include the attachment of proteins,oligonucleotides, drugs, ligands, catalysts, dyes, sensors, analytes,and charged species capable of changing the wettability of the channel.

V.C. Method of Linking Multiple Chains of a PFPE Material with aFunctional Linker Group

In some embodiments, the presently disclosed method adds functionalityto a microfluidic channel or a microtiter well by adding a chemical“linker” moiety to the elastomer itself. In some embodiments, afunctional group is added along the backbone of the precursor material.An example of this method is illustrated in Scheme 8.

In some embodiments, the precursor material includes a macromoleculecontaining hydroxyl functional groups. In some embodiments, as depictedin Scheme 8, the hydroxyl functional groups include diol functionalgroups. In some embodiments, two or more of the diol functional groupsare connected through a trifunctional “linker” molecule. In someembodiments, the trifunctional linker molecule has two functionalgroups, R and R′. In some embodiments, the R′ group reacts with thehydroxyl groups of the macromolecule. In Scheme 8, the circle canrepresent a linking molecule; and the wavy line can represent a PFPEchain.

In some embodiments, the R group provides the desired functionality tothe interior surface of the microfluidic channel or surface of amicrotiter well. In some embodiments, the R′ group is selected from thegroup including, but not limited to, an acid chloride, an isocyanate, ahalogen, and an ester moiety. In some embodiments, the R group isselected from one of, but not limited to, a protected amine and aprotected alcohol. In some embodiments, the macromolecule diol isfunctionalized with polymerizable methacrylate groups. In someembodiments, the functionalized macromolecule diol is cured and/ormolded by a photochemical process as described by Rolland, J. et al.JACS 2004, 126, 2322-2323, the disclosure of which is incorporatedherein by reference in its entirety.

Thus, the presently disclosed subject matter provides a method ofincorporating latent functional groups into a photocurable PFPE materialthrough a functional linker group. Thus, in some embodiments, multiplechains of a PFPE material are linked together before encapping the chainwith a polymerizable group. In some embodiments, the polymerizable groupis selected from the group consisting of a methacrylate, an acrylate,and a styrenic. In some embodiments, latent functionalities are attachedchemically to such “linker” molecules in such a way that they will bepresent in the fully cured network.

In some embodiments, latent functionalities introduced in this mannerare used to bond multiple layers of PFPE, bond a fully cured PFPE layerto a substrate, such as a glass material or a silicon material that hasbeen treated with a silane coupling agent, or bond a fully cured PFPElayer to a second polymeric material, such as a PDMS material. In someembodiments, the PDMS material is treated with a plasma and a silanecoupling agent to introduce the desired functionality. In someembodiments, the PDMS material is encapped with a polymerizable group.In some embodiments, the polymerizable group is selected from the groupconsisting of an acrylate, a styrene, and a methacrylate.

In some embodiments, the second polymeric material includes an elastomerother than PDMS, such as Kratons™, buna rubber, natural rubber, afluoroelastomer, chloroprene, butyl rubber, nitrile rubber,polyurethane, or a thermoplastic elastomer. In some embodiments, thesecond polymeric material includes a rigid thermoplastic, including butnot limited to: polystyrene, poly(methyl methacrylate), a polyester,such as poly(ethylene terephthalate), a polycarbonate, a polyimide, apolyamide, a polyvinylchloride, a polyolefin, a poly(ketone), apoly(ether ether ketone), and a poly(ether sulfone).

In some embodiments, PFPE networks including functionality attached to“linker” molecules are used to functionalize the interior surface of amicrofluidic channel and/or the surface of a microtiter well. In someembodiments, the inside of a microfluidic channel is functionalized byattaching a functional moiety selected from the group consisting of aprotein, an oligonucleotide, a drug, a catalyst, a dye, a sensor, ananalyte, and a charged species capable of changing the wettability ofthe channel.

V.D. Methods for Improving Chemical Compatibility of a Surface

According to some embodiments of the present invention, the surface ofdevices fabricated from materials and methods described herein can bepassivated to impart chemical compatibility to the devices. According tosuch materials and methods, surface passivation is achieved by treatingthe surface of a device fabricated from materials described herein withan end-capped UV and/or thermal curable liquid precursor (e.g., styreneend-capped precursor). Upon activation of the photo or thermally curecomponent of the styrene end-capped precursor, the precursor reacts withlatent methacrylate, styrene, and/or acrylate groups of the material andbinds thereto, thereby providing a surface passivation to the surface ofthe device.

According to another embodiment, a device fabricated from PFPE thatcontains latent methacrylate, acrylate, and/or styrene groups, asdescribed throughout this application, is treated with a styreneend-capped UV curable PFPE liquid precursor. According to suchembodiments, a solution of the styrene end-capped UV curable precursor,dissolved in a solvent including but not limited to pentafluorobutane,can be applied to a surface of a device fabricated from: PFPE. Thesolvent is allowed to evaporate, thereby leaving a film of the styreneend-capped UV curable precursor coating the PFPE surface. In oneembodiment the film is then cured, by exposure to UV light, and therebyadhered to latent methacrylate, acrylate, and/or styrene groups of thePFPE material. The surface coated with the styrene end-capped precursordoes not contain acid-labile groups such as urethane and/or esterlinkages, thus creating a surface passivation and improving the chemicalcompatibility of the base PFPE material.

According to another embodiment, the surface of a device fabricated frombase materials described herein is passivated by a gas phasepassivation. According to such embodiments, a device is exposed to amixture of 0.5% Fluorine gas in Nitrogen. The Fluorine reacts freeradically with hydrogen atoms in the base material, thus passivating thesurface of device that is treated with the gas.

VI. Method of Adding Functional Monomers to the PFPE Precursor Material

In some embodiments, the method includes adding a functional monomer toan uncured precursor material. In some embodiments, the functionalmonomer is selected from the group consisting of functional styrenes,methacrylates, and acrylates. In some embodiments, the precursormaterial includes a fluoropolymer. In some embodiments, the functionalmonomer includes a highly fluorinated monomer. In some embodiments, thehighly fluorinated monomer includes perfluoro ethyl vinyl ether (EVE).In some embodiments, the precursor material includes a poly(dimethylsiloxane) (PDMS) elastomer. In some embodiments, the precursor materialincludes a polyurethane elastomer. In some embodiments, the methodfurther includes incorporating the functional monomer into the networkby a curing step.

In some embodiments, functional monomers are added directly to theliquid PFPE precursor to be incorporated into the network uponcrosslinking. For example, monomers can be introduced into the networkthat are capable of reacting post-crosslinking to adhere multiple layersof PFPE, bond a fully cured PFPE layer to a substrate, such as a glassmaterial or a silicon material that has been treated with a silanecoupling agent, or bond a fully cured PFPE layer to a second polymericmaterial, such as a PDMS material. In some embodiments, the PDMSmaterial is treated with a plasma and a silane coupling agent tointroduce the desired functionality. In some embodiment, the PDMSmaterial is encapped with a polymerizable group. In some embodiments,the polymerizable material is selected from the group consisting of anacrylate, a styrene, and a methacrylate.

In some embodiments, the second polymeric material includes an elastomerother than PDMS, such as Kratons™, buna rubber, natural rubber, afluoroelastomer, chloroprene, butyl rubber, nitrile rubber,polyurethane, or a thermoplastic elastomer. In some embodiments, thesecond polymeric material includes a rigid thermoplastic, including butnot limited to: polystyrene, poly(methyl methacrylate), a polyester,such as poly(ethylene terephthalate), a polycarbonate, a polyimide, apolyamide, a polyvinylchloride, a polyolefin, a poly(ketone), apoly(ether ether ketone), and a poly(ether sulfone).

In some embodiments, functional monomers are added directly to theliquid PFPE precursor and are used to attach a functional moietyselected from the group consisting of a protein, an oligonucleotide, adrug, a catalyst, a dye, a sensor, an analyte, and a charged speciescapable of changing the wettability of the channel.

Such monomers include, but are not limited to, tert-butyl methacrylate,tert butyl acrylate, dimethylaminopropyl methacrylate, glycidylmethacrylate, hydroxy ethyl methacrylate, aminopropyl methacrylate,allyl acrylate, cyano acrylates, cyano methacrylates, trimethoxysilaneacrylates, trimethoxysilane methacrylates, isocyanato methacrylate,lactone-containing acrylates and methacrylates, sugar-containingacrylates and methacrylates, poly-ethylene glycol methacrylate,nornornane-containing methacrylates and acrylates, polyhedral oligomericsilsesquioxane methacrylate, 2-trimethylsiloxyethyl methacrylate,1H,1H,2H,2H-fluoroctylmethacrylate, pentafluorostyrene, vinyl pyridine,bromostyrene, chlorostyrene, styrene sulfonic acid, fluorostyrene,styrene acetate, acrylamide, and acrylonitrile.

In some embodiments, monomers which already have the above agentsattached are blended directly with the liquid PFPE precursor to beincorporated into the network upon crosslinking. In some embodiments,the monomer includes a group selected from the group consisting of apolymerizable group, the desired agent, and a fluorinated segment toallow for miscibility with the PFPE liquid precursor. In someembodiments, the monomer does not include a polymerizable group, thedesired agent, and a fluorinated segment to allow for miscibility withthe PFPE liquid precursor.

In some embodiments, monomers are added to adjust the mechanicalproperties of the fully cured elastomer. Such monomers include, but arenot limited to: perfluoro(2,2-dimethyl-1,3-dioxole), hydrogen-bondingmonomers which contain hydroxyl, urethane, urea, or other such moieties,monomers containing bulky side group, such as tert-butyl methacrylate.

In some embodiments, functional species such as the above mentionedmonomers are introduced and are mechanically entangled, i.e., notcovalently bonded, into the network upon curing. For example, in someembodiments, functionalities are introduced to a PFPE chain that doesnot contain a polymerizable monomer and such a monomer is blended withthe curable PFPE species. In some embodiments, such entangled speciescan be used to adhere multiple layers of cured PFPE together if twospecies are reactive, such as: epoxy/amine, hydroxy/acid chloride,triol/diisocyanate, hydroxy/isocyanate, amine/isocyanate, amine/halide,hydroxy/halide, amine/ester, and amine/carboxylic acid. Upon heating,the functional groups will react and adhere the two layers together.

Additionally, such entangled species can be used to adhere a PFPE layerto a layer of another material, such as glass, silicon, quartz, PDMS,Kratons™, buna rubber, natural rubber, a fluoroelastomer, chloroprene,butyl rubber, nitrile rubber, polyurethane, or a thermoplasticelastomer. In some embodiments, the second polymeric material includes arigid thermoplastic, including but not limited to: polystyrene,poly(methyl methacrylate), a polyester, such as poly(ethyleneterephthalate), a polycarbonate, a polyimide, a polyamide, apolyvinylchloride, a polyolefin, a poly(ketone), a poly(ether etherketone), and a poly(ether sulfone).

In some embodiments, such an entangled species can be used tofunctionalize the interior of a microfluidic channel for the purposesdescribed hereinabove.

VII. Other Methods of Introducing Functionality to a PFPE Surface

In some embodiments, an Argon plasma is used to introduce functionalityalong a fully cured PFPE surface using the method for functionalizing apoly(tetrafluoroethylene) surface as described by Chen, Y. and Momose,Y. Surf Interface. Anal. 1999, 27, 1073-1083, which is incorporatedherein by reference in it entirety. More particularly, without beingbound to any one particular theory, exposure of a fully cured PFPEmaterial to Argon plasma for a period of time adds functionality alongthe fluorinated backbone.

Such functionality can be used to adhere multiple layers of PFPE, bond afully cured PFPE layer to a substrate, such as a glass material or asilicon material that has been treated with a silane coupling agent, orbond a fully cured PFPE layer to a second polymeric material, such as aPDMS material. In some embodiments, the PDMS material includes afunctionalized material. In some embodiments, the PDMS material istreated with a plasma and a silane coupling agent to introduce thedesired functionality. Such functionalities also can be used to attachproteins, oligonucleotides, drugs, catalysts, dyes, sensors, analytes,and charged species capable of changing the wettability of the channel.

In some embodiments, the second polymeric material includes an elastomerother than PDMS, such as Kratons™, buna rubber, natural rubber, afluoroelastomer, chloroprene, butyl rubber, nitrile rubber,polyurethane, or a thermoplastic elastomer. In some embodiments, thesecond polymeric material includes a rigid thermoplastic, including butnot limited to: polystyrene, poly(methyl methacrylate), a polyester,such as poly(ethylene terephthalate), a polycarbonate, a polyimide, apolyamide, a polyvinylchloride, a polyolefin, a poly(ketone), apoly(ether ether ketone), and a poly(ether sulfone).

In some embodiments, a fully cured PFPE layer is brought into conformalcontact with a solid substrate. In some embodiments, the solid substrateis selected from the group consisting of a glass material, a quartzmaterial, a silicon material, a fused silica material, and a plasticmaterial. In some embodiments, the PFPE material is irradiated with UVlight, e.g., a 185-nm UV light, which can strip a fluorine atom off ofthe back bone and form a chemical bond to the substrate as described byVurens, G., et al. Langmuir 1992, 8, 1165-1169. Thus, in someembodiments, the PFPE layer is covalently bonded to the solid substrateby radical coupling following abstraction of a fluorine atom.

VIII. Adhesion of a Microscale or a Nanoscale Device to a SubstrateThrough an Encasing Polymer

In some embodiments, a microscale device, a nanoscale device, orcombinations thereof is adhered to a substrate by placing the fullycured device in conformal contact on the substrate and pouring an“encasing polymer” over the entire device. In some embodiments, theencasing polymer is selected from the group consisting of a liquid epoxyprecursor and a polyurethane. The encasing polymer is then solidified bycuring or other methods. The encasement serves to bind the layerstogether mechanically and to bind the layers to the substrate.

In some embodiments, the microscale device, the nanoscale device, orcombinations thereof includes one of a perfluoropolyether material asdescribed in Section II.A and Section II.B., hereinabove, and afluoroolefin-based material as described in Section II.C., hereinabove.

In some embodiments, the substrate is selected from the group consistingof a glass material, a quartz material, a silicon material, a fusedsilica material, and a plastic material. Further, in some embodiments,the substrate includes a second polymeric material, such aspoly(dimethylsiloxane) (PDMS), or another polymer. In some embodiments,the second polymeric material includes an elastomer other than PDMS,such as Kratons™, buna rubber, natural rubber, a fluoroelastomer,chloroprene, butyl rubber, nitrile rubber, polyurethane, or athermoplastic elastomer. In some embodiments, the second polymericmaterial includes a rigid thermoplastic material, including but notlimited to: polystyrene, poly(methyl methacrylate), a polyester, such aspoly(ethylene terephthalate), a polycarbonate, a polyimide, a polyamide,a polyvinylchloride, a polyolefin, a poly(ketone), a poly(ether etherketone), and a poly(ether sulfone). In some embodiments, the surface ofthe substrate is functionalized with a silane coupling agent such thatit will react with the encasing polymer to form an irreversible bond.

IX. Method for Forming a Microstructure Using Sacrificial Layers

The presently disclosed subject matter provides a method for formingmicrochannels or a microstructure for use as a microfluidic device byusing sacrificial layers including a degradable or selectively solublematerial. In some embodiments, the method includes contacting a liquidprecursor material with a two-dimensional or a three-dimensionalsacrificial structure, treating, e.g., curing, the precursor material,and removing the sacrificial structure to form a microfluidic channel.

Accordingly, in some embodiments, a PFPE liquid precursor is disposed ona multidimensional scaffold, wherein the multidimensional scaffold isfabricated from a material that can be degraded or washed away aftercuring of the PFPE network. These materials protect the channels frombeing filled in when another layer of elastomer is cast thereon.Examples of such degradable or selective soluble materials include, butare not limited to waxes, photoresists, polysulfones, polylactones,cellulose fibers, salts, or any solid organic or inorganic compounds. Insome embodiments, the sacrificial layer is removed thermally,photochemically, or by washing with solvents. Importantly, thecompatibility of the materials and devices disclosed herein with organicsolvents provides the capability to use sacrificial polymer structuresin microfluidic devices.

The PFPE materials of use in forming a microstructure by usingsacrificial layers include those PFPE and fluoroolefin-based materialsas described hereinabove in Section II of the presently disclosedsubject matter.

FIGS. 6A-6D and FIGS. 7A-7C show embodiments of the presently disclosedmethods for forming a microstructure by using a sacrificial layer of adegradable or selectively soluble material.

Referring now to FIG. 6A, a patterned substrate 600 is provided. LiquidPFPE precursor material 602 is disposed on patterned substrate 600. Insome embodiments, liquid PFPE precursor material 602 is disposed onpatterned substrate 600 via a spin-coating process. Liquid PFPEprecursor material 602 is treated by treating process T_(r1) to form alayer of treated liquid PFPE precursor material 604.

Referring now to FIG. 6B, the layer of treated liquid PFPE precursormaterial 604 is removed from patterned substrate 600. In someembodiments, the layer of treated liquid PFPE precursor material 604 iscontacted with substrate 606. In some embodiments, substrate 606includes a planar substrate or a substantially planar substrate. In someembodiments, the layer of treated liquid PFPE precursor material istreated by treating process T_(r2), to form two-layer assembly 608.

Referring now to FIG. 6C, a predetermined volume of degradable orselectively soluble material 610 is disposed on two-layer assembly 608.In some embodiments, the predetermined volume of degradable orselectively soluble material 610 is disposed on two-layer assembly 608via a spin-coating process. Referring once again to FIG. 6C, liquidprecursor material 602 is disposed on two-layer assembly 608 and treatedto form a layer of PFPE material 612, which covers the predeterminedvolume of degradable or selectively soluble material 610.

Referring now to FIG. 6D, the predetermined volume of degradable orselectively soluble material 610 is treated by treating process T_(r3)to remove the predetermined volume of degradable or selectively solublematerial 610, thereby forming microstructure 616. In some embodiments,microstructure 616 includes a microfluidic channel. In some embodiments,treating process T_(r3) is selected from the group consisting of athermal process, an irradiation process, and a dissolution process.

In some embodiments, patterned substrate 600 includes an etched siliconwafer. In some embodiments, the patterned substrate includes aphotoresist patterned substrate. For the purposes of the presentlydisclosed subject matter, the patterned substrate can be fabricated byany of the processing methods known in the art, including, but notlimited to, photolithography, electron beam lithography, and ionmilling.

In some embodiments, degradable or selectively soluble material 610 isselected from the group consisting of a polyolefin sulfone, a cellulosefiber, a polylactone, and a polyelectrolyte. In some embodiments, thedegradable or selectively soluble material 610 is selected from amaterial that can be degraded or dissolved away. In some embodiments,degradable or selectively soluble material 610 is selected from thegroup consisting of a salt, a water-soluble polymer, and asolvent-soluble polymer.

In addition to simple channels, the presently disclosed subject matteralso provides for the fabrication of multiple complex structures thatcan be “injection molded” or fabricated ahead of time and embedded intothe material and removed as described above.

FIGS. 7A-C illustrate an embodiment of the presently disclosed methodfor forming a microchannel or a microstructure through the use of asacrificial layer. Referring now to FIG. 7A, a substrate 700 isprovided. In some embodiments, substrate 700 is coated with a liquidPFPE precursor material 702. Sacrificial structure 704 is placed onsubstrate 700. In some embodiments, liquid PFPE precursor material 702is treated by treating process T_(r1).

Referring now to FIG. 7B, a second liquid PFPE precursor material 706 isdisposed over sacrificial structure 704, in such a way to encasesacrificial structure 704 in second liquid precursor material 706.Second liquid precursor material 706 is then treated by treating processT_(r2). Referring now to FIG. 7C, sacrificial structure 704 is treatedby treating process T_(r3), to degrade and/or remove sacrificialstructure, thereby forming microstructure 708. In some embodiments,microstructure 708 includes a microfluidic channel.

In some embodiments, substrate 700 includes a silicon wafer. In someembodiments, sacrificial structure 704 includes a degradable orselectively soluble material. In some embodiments, sacrificial structure704 is selected from the group consisting of a polyolefin sulfone, acellulose fiber, a polylactone, and a polyelectrolyte. In someembodiments, the sacrificial structure 704 is selected from a materialthat can be degraded or dissolved away. In some embodiments, sacrificialstructure 704 is selected from the group consisting of a salt, awater-soluble polymer, and a solvent-soluble polymer.

IX.I. Method of Increasing the Modulus of a Microfluidic Device UsingPTFE Powder

In some embodiments, the modulus of a microfluidic device fabricatedfrom PFPE materials or any of the fluoropolymer materials describedhereinabove can be increased by blending polytetrafluoroethylene (PTFE)powder, also referred to herein as a “PTFE filler,” into the liquidprecursor prior to curing. Because PTFE itself has a very high modulus,addition of PTFE in its powder form, when evenly dispersed throughoutthe low modulus materials of the present invention, will raise theoverall modulus of the material. The PTFE filler also can contributeadditional chemical stability and solvent resistance to the PFPEmaterials.

IX.II. Use of the Material in Combination with a Microfluidic Device

According to an embodiment of the present invention, micro or nano scaledevices, portions of devices, components, parts, or the like made fromthe methods and materials described herein can be formed forincorporation into a microfluidic device. For example, micro or nanoscale valves or plugs can be formed from the materials and methods ofthe present invention that can effectively close off channels in amicrofluidic device. According to one embodiment, the valve or plug canbe formed in a shape and/or size configuration to fit within amicro-chamber and remain in position or be configured to move inresponse to substances flowing in a particular direction or blockparticular channels from flow. According to another embodiment, a valveor plug can be formed in an micro-channel by introducing the materialsof the present invention, in liquid form, into the micro-channel andcuring the liquid material according to the methods disclosed in thepresent invention. Thereby, the valve or plug takes on the shape of themicro-channel forming a conformal fit because the channel acts as themold for the valve. In alternative embodiments, a valve is formed bypositioning a first channel adjacent a test channel such that when thefirst channel is pressurized the first channel exerts a pressure on thetest channel and reduces a cross-section width of the test channel,thereby reducing or eliminating the flow path within the test channel.

X. Microfluidic Devices

X.A. Functionalizing Microfluidic Devices

According to embodiments of the present invention microfluidic devicescan be functionalized to increase their chemical compatibility.According to such embodiments the surface of channels of themicrofluidic devices can be passivated by methods and materials of thepresent invention. According to such embodiments, a PFPE basedmicrofluidic device can be treated with a styrene end-capped UV curableliquid precursor. The styrene end-capped UV curable precursor isdissolved in a solvent, such as but not limited to, pentafluorobutanesuch that a solution is formed. This solution is then introduced intothe channels of the microfluidic device and the solvent is evaporated.Following evaporation of the solvent, the precursor remains coated onthe walls of the channels. In one embodiment, the microfluidic devicewith the precursor coated on the walls of the channels is treated with aUV treatment which adheres the precursor to the walls of the channelsthrough reaction with latent methacrylate groups contained in the basematerial of the microfluidic device. The surface coated with the styreneend-capped precursor does not contain acid-labile groups such asurethane and/or ester linkages, thus creating a surface passivation andimproving the chemical compatibility of the channels of the microfluidicdevice.

According to other embodiments, medical devices, surgical devices,medical implants, and the like, fabricated from materials and methodsdisclosed in the present application can be treated with the surfacetreatment methods herein described to create surface passivation on thedevice and increase chemical compatibility of the devices. Such surfacepassivation can increase the chemically inert nature of the basematerial, reduce adsorption of substances to the surface, increaseresistance to acids and bases, combinations thereof, and the like.

According to another embodiment, the surface of a device fabricated frommaterials described herein is passivated by a gas phase passivation.According to such embodiments, a device is exposed to a mixture of 0.5%Fluorine gas in Nitrogen. The Fluorine reacts free radically withhydrogen atoms in the base material, thus passivating the surface ofdevice that is treated with the gas.

X.B. Devices Having Torque Actuated Valves

In some embodiments, microfluidic devices are fabricated from thematerials and methods described herein and contain “torque actuatedvalves” such as those described by Whitesides et al. in Anal Chem 2005,77, 4726. Such valves, as well as the valves described in the referencesbelow are incorporated into fluoropolymer or PFPE-based microfluidicchips. References describing appropriate valves that can be fabricated,treated, utilized, or the like from the materials and methods disclosedherein include: (1) Lee, S.; Jeong, W.; Beebe, D. J. Lab Chip 2003, 3,164-167; (2) Rich, C. A.; Wise, K. D. J. Microelectromech. Syst. 2003,12, 201-208; (3) Studer, V.; Hang, G.; Pandolfi, A.; Ortiz, M.;Anderson, W. F.; Quake, S. R; J. Appl. Phys. 2004, 95, 393-398; (4) Sin,A.; Reardon, C. F.; Shuler, M. L. Biotechnol. Bioeng. 2004, 85, 359-363;(5) Gu, W.; Zhu, X.; Futai, N.; Cho, B. S.; Takayama, S. Proc. Natl.Acad. Sci. U.S.A. 2004, 101, 15861-15866; (6) Rohit, P.; Yang, M.;Johnson, B. N.; Burns, D. T.; Burns, M. A. Anal. Chem. 2004, 76,3740-3748; (7) Liu, R. H.; Bonanno, J.; Yang, J.; Lenigk, R.;Grodzinski, P. Sens. Actuators, B 2004, B98, 328-336; (8)Selvaganaphthy, P.; Carlen, E. T.; Mastrangelo, C. H. Sens. Actuators, A2003, A104, 275-282; (9) Sethu, P.; Mastrangelo, C. H. Sens. Actuators,A 2003, A104, 283-289; (10) Klintberg, L.; Svedberg, M.; Nikolajeff, F.;Thornell, G. Sens. Actuators, A 2003, A103, 307-316; (11) Suzuki, H.;Yoneyama, R. Sens. Actuators, B 2003, B96, 38-45; (12) Hua, S. Z.;Sachs, F.; Yang, D. X, Chopra, H. D. Anal. Chem. 2002, 74, 6392-6396;(13) Xie, J.; Miao, Y.; Shih, J.; He, Q.; Liu, J.; Tai, Y.-C.; Lee, T.D. Anal. Chem. 2004, 76, 3756-3763; (14) Tsai, J. H.; Lin, L. J.Microelectromech. Syst. 2002, 11, 665-671; (15) Munyan, J. W.; Fuentes,H. V.; Draper, M.; Kelly, R. T.; Woolley, A. T. Lab Chip 2003, 3,217-220; (16) Hartshorne, H.; Backhouse, C. J.; Lee, W. E. Sens.Actuators, B 2004, B99, 592-600; (17) Hatch, A.; Kamholz, A. E.; Holman,G.; Yager, P.; Bohringer, K. F. J. Microelectromech. Syst. 2001, 10,215-221; (18) Jackson, W. C.; Tran, H. D.; O'Brien, M. J.; Rabinovich,E.; Lopez, G. P. J. Vac. Sci. Technol. B 2001, 19, 596-599; (19) Luo,Q.; Mutlu, S.; Gianchandani, Y. B.; Svec, F.; Frechet, J. M. J.Electrophoresis 2003, 24, 3694-3702; (20) Yu, C.; Mutlu, S.;Selvaganaphthy, P.; Mastrangelo, C. H.; Svec, F.; Frechet, J. M. J.Anal. Chem. 2003, 75, 1958-1961; (21) Griss, P.; Andersson, H.; Stemme,G. Lab Chip 2002, 2, 117-120; (22) Beebe, D. J.; Moore, J. S.; Bauer, J.M.; Yu, Q.; Liu, R. H.; Devadoss, C.; Jo, B. Nature 2000, 404, 588-590;(23) Beebe, D. J.; Mensing, G. A.; Walker, G. M. Annu. Rev. Biomed. Eng.2002 4, 261-286; (24) Jacobson, S. C.; Ermakov, S. V.; Ramsey, J. M.Anal. Chem. 1999, 71, 3273-3276; (25) Lazar, I. M.; Karger, B. L. Anal.Chem. 2002, 74, 6259-6268; (26) Gitlin, I.; Stroock, A. D.; Whitesides,G. M.; Ajdari, A. Appl. Phys. Lett. 2003, 83, 1486-1488; (27)Lastochkin, D.; Zhou, R.; Wang, P.; Ben, Y.; Chang, H.-C. J. Appl. Phys.2004, 96, 1730-1733; (28) Liu, S.; Pu, Q.; Lu, J. J. J. Chromatogr., A2003, 1013, 57-64; (29) Takamura, Y.; Onoda, H.; Inokuchi, H.; Adachi,S.; Oki, A.; Horiike, Y. Electrophoresis 2003, 24, 185-192; (30)McKnight, T. E.; Culbertson, C. T.; Jacobson, S. C.; Ramsey, J. M. AnalChem. 2001, 73, 4045-4049; (31) Culbertson, C. T.; Ramsey, R. S.;Ramsey, M. J. Anal. Chem. 2000, 72, 2285-2291; (32) Salimi-Moosavi, H.;Tang, T.; Harrison, D. J. J. Am. Chem. Soc. 1997, 119, 8716-8717; (33)Johnston, I. D.; Davis, J. B.; Richter, R.; Herbert, G. I.; Tracey, M.C. Analyst 2004, 129, 829-834; (34) Terray, A.; Oakey, J.; Marr, D. W.M. Science 2002, 296, 1841-1844; (35) Andersson, H.; van der Wijngaart,W.; Griss, P.; Niklaus, F.; Stemme, G. Sens. Actuators, B 2001, B75,136-141; (36) Walker, G. M.; Beebe, D. J. Lab Chip 2002, 2, 131-134;(37) Shoji, S. Top. Curr. Chem. 1998, 194, 163-188; (38) Kovacs, G. T.A. Micromachined Transducers Sourcebook; McGraw-Hill: New York, 1998;each of which is incorporated herein by reference in its entirety.

X.C. Multiple Material Devices

In some embodiments, the materials described herein are used in only apart of a fabricated microfluidic device. In such devices, portions ofthe device are made from the materials disclosed herein and otherportions of the device are made from other materials described hereinsuch as glass or PDMS. Examples of such “parts” include but are notlimited to: valves, channels, walls, discs, layers, backing, plugs,balls, switches, gears, and pillars. In some embodiments, thesestructures can be generated in situ by exposing UV light through aphotomask of the desired shape such that the PFPE material is cured inthe desired shape in select regions. Residual PFPE material that was notexposed to UV light due to the mask are left untreated and can then bewashed away with a solvent such as 1,1,1-3,3-pentafluorobutane. Thisleaves a structure behind that mimics the mask opening. In furtherembodiments, this structure is adhered in place by activating a secondcuring step as described previously.

X.D. Master Template Patterns for Fabricating Devices

Referring to FIG. 17, in some embodiments, master template patterns usedto fabricate devices, such as for example microfluidic devices, can betransferred to a PDMS slab which is then used to pattern devices.According to such embodiments, an initial master template 1702 isprepared with a pattern 1704 that is to be transferred to a finaldevice. In some embodiments master template 1702 can be generated from asilicon substance, a photoresist substance, combinations thereof, or thelike. Next, a liquid PFPE precursor 1706 is introduced to mastertemplate 1702 and/or its pattern 1704, as shown in steps B-C. In someembodiments, liquid PFPE 1706 can be introduced onto master template1702 by casting, deposition, spin coating, pressing, pouring,combinations thereof, or the like. Liquid PFPE 1706 is then cured whilein contact with master template 1702 to form solidified liquid PFPE 1708that can retain pattern 1704 of master template 1702 when removed frommaster template 1702. In some embodiments, liquid PFPE 1704 is cured asdescribed herein, such as for example by, photo-curing, thermal curing,evaporation curing, combinations thereof, and the like. Solidifiedliquid PFPE 1708 is then removed from master template 1702, as shown instep C. Solidified liquid PFPE 1708 can then be used as a template forpatterning a PDMS layer 1710, as shown in steps D-F. Liquid PDMS 1710 isbrought into contact with solidified liquid PFPE 1708 and cured, formingsolidified liquid PDMS 1712, as shown in steps E-F. Liquid PDMS 1710 caninclude curing agents as described herein and can be cured by methodsand techniques described herein, such as for example, photo activationof photo-curing agents, thermal activation of thermal curing agents,evaporation curing, combinations thereof, and the like. Solidifiedliquid PDMS 1712 is then removed from solidified liquid PFPE template1708, as shown in step F. Finally, solidified liquid PDMS 1712 can beused as a template to mold subsequent PFPE devices by introducing asecond quantity of liquid PFPE 1714 to solidified liquid PDMS template1712 and curing liquid PFPE 1714, as shown in steps G-H. Liquid PFPE1714 can include curing agents as described herein and can be cured bymethods and techniques described herein to form PFPE device 1716. PFPEdevice 1716 can then be removed from solidified liquid PDMS template1710. The use of liquid PDMS as a template for fabrication of PFPEdevice 1716 allows for easier release of PFPE device 1716 from the PDMStemplate 1710 than release from a silicon or photoresist template. Useof a PDMS template, such as described herein, if further advantageousover a silicon or photoresist template when the silicon or photoresisttemplate contains latent functionalities which can react with theendgroups on the PFPE precursors, thereby causing adhesion of the PFPEdevice to the silicon or photoresist template.

X.E. Layering by Spin Coating

In some embodiments, layers of materials described herein can be adheredto other layers by spin coating a thin layer of uncured material ontoanother layer, positioning the layers with respect to each other, andcuring the material, with for example UV light, photo-curing, or thermalenergy, to activate curing agents within the layers and/or spin coatedthin layer such that the layers are adhered together. In someembodiments this process is repeated multiple times to form a multilayerdevice. In some embodiments the spin coated thin layer can be betweenabout 50 nm and about 2 microns. According to other embodiments, thethin layer is between about 100 nm and about 1 micron.

X.F. Low Permeability Layer Inside a Device

In some embodiments, a layer of a material with very low permeability toa liquid or gas such as oxygen, water vapor, water, combinationsthereof, or the like is fabricated as an inner layer of a multiple layerdevice. Such device can be, for example, a microfluidic device asdescribed herein. The low permeability material can be, for example, anacrylic polymer such as poly(methyl methacrylate), poly(ethylene), or aUV curable resin such as trimethylolpropane triacrylate, or a materialmade by metathesis polymerization such as poly(dicyclo pentadiene),combinations thereof, or the like.

X.G. Acid and/or Base Treatment

In some embodiments, a layer of materials described herein can betreated with an acid and/or base such that any latent functionalitiesassociated with a surface of the material can be neutralized.Neutralizing such latent functionalities reduces interactions withsubstances that come into contact with the material during use.According to some embodiments, the surface of the material treated isthe surface of a channel formed within the material, such as forexample, a channel of a microfluidic device. According to someembodiments, F— is a preferable surface treatment, however othertreatments can also be utilized and may be more appropriate for thecompositions of the material to be treated.

X.H. Microfluidic Device with Minimum Quenching of Reactions InvolvingF—

According to some embodiments of the present invention, microfluidicdevices contain from substantially zero to minimal amounts of tracemetals. In some embodiments, the microfluidic devices are fabricatedfrom PFPE materials that contain from substantially zero to minimalamounts of trace metals.

Many reactions involving F— as a nucleophile are quenched by traceamounts of metal catalysts, specifically, organo-tin catalysts such asdibutyltin diacetate, dibutyltin dilaurate, and stannous octoate.Further, it is generally known that metal catalysts or metal complexeswill form strong complexes with F—. For example, metal complexescontaining Ru, Rh, Pd, Pt, Cu, Co, Ni, Fe, Cr, Sn, Al, and Si are knownto form strong associations with F—. An example of such a reaction isshown below for the synthesis of 2-deoxy-2-[¹⁸F]fluoro-D-glucose (FDG),an imaging agent used in positron emission tomography (PET).

Many reactions using microfluidic devices involve the synthesis of PETagents. Moreover, many if not most of these agents involve reactionsusing F— as a nucleophile. Therefore, microfluidic device fabricatedfrom the materials disclosed herein are preferable for PET reactionsusing F— as a nucleophile because.

X.I. Use of an Epoxy-functional Polymer Containing a Photoacid Generator

In some embodiments, a layer of an epoxy-functional polymer containing aphotoacid generator can be deposited onto a surface or in betweenmultiple layers of a device. In some embodiments, the epoxy functionalpolymer has a Tg at or above room temperature and is spin-coated onto asurface such as glass, silicon, a flat or cured layer of PFPE material,combinations thereof, or the like. Layers of a device can then be sealedto the polymer layer and irradiated with UV light. The UV-lightactivates the PAG within the epoxy-functional polymer causing it tocrosslink as well as react with latent groups on the surface of the PFPElayer, thus adhering the PFPE layer to the surface. In some embodiments,the epoxy-functional polymer consists of SU-8 photoresist, polymerscontaining the following structure, combinations thereof, or the like:

II.J. Epoxy-functional Polymers

In some embodiments, the epoxy-functional polymer, as described hereincan be a copolymer of glycidyl methacrylate and one or more comonomerssuch as Zonyl TM® fluoroalkyl methacrylate monomers or the like.

II.K. ePTFE Materials in Combination with PFPE

In some embodiments, a microfluidic device or a component of amicrofluidic device (e.g., membrane, valve, channel, reservoir, or thelike) is formed by using expanded poly(tetrafluoroethylene) (ePTFE).ePTFE is a microporous poly(tetrafluoroethylene) (PTFE) membrane formedby expanding PTFE at high temperatures. Because of the incredibly lowsurface energy of perfluoropolyether (PFPE) materials, as disclosedherein, the PFPE can be cast onto ePTFE membranes and effectively wetthe pores included in the ePTFE, thereby forming an interpenetratingpolymer network.

The PFPE can include functional groups, such as functional groupsdisclosed herein. In some embodiments, the PFPE can include photocurablefunctional groups and/or thermalcurable functional groups. In someembodiments, where the PFPE includes a photocurable functional group,the PFPE can be cured by exposure to UV light after it has been wettedonto the ePTFE. The resulting membrane of ePTFE and PFPE includes, forexample, desirable flexibility, chemically resistance, and gaspermeability to solvent vapors and to air. The ePTFE portion of themembrane provides a continuous structure which serves to greatly toughenmaterials compared to just an elastomer itself. Some membranes aregenerally described by Zumbrum et al., in U.S. Pat. No. 6,673,455, whichis incorporated by reference herein in its entirety including allreference cited therein.

In some embodiments, membranes of combined ePTFE and PFPE can be used inmicrofluidic platforms, especially where evaporation of solvents througha membrane is desired. According to some embodiments, these membranesare useful in microfluidic platforms involving the synthesis ofradiolabelled biomarkers for Positron Emission Tomography (PET) imagingwhere chemical inertness of the membrane can be crucial. In someembodiments, the ePTFE membrane can be filled with a PFPE distyrenematerial that can be subsequently UV or thermally cured, yielding highchemical resistance.

In other embodiments, the “dual cure” PFPE materials described hereincan be used to wet ePTFE membranes or microfluidic device components andactivated in such ways to adhere the membrane to other membranes orother components of the device. In some embodiments, methods forfabricating a microfluidic device (or device in general) can includetaking a mixture of UV curable and thermally curable PFPE materials andcasting them onto an ePTFE membrane, UV curing the PFPE, sealing thecombination of ePTFE/PFPE to another layer or component of amicrofluidic device, and then heating the combination of ePTFE/PFPE anddevice component to activate the thermal component, thus generatingadhesion of the device component to the PFPE layer that is adhered tothe ePTFE.

XI. Microfluidics Unit Operations

Microfluidic control devices are necessary for the development ofeffective lab-on-a-chip operations. Valve structures and actuation,fluid control, mixing, separation, and detection at microscale levelsmust be designed to have a large-scale shift to miniaturization. Toconstruct such devices, integration of the individual components on acommon platform must be developed so that solvents and solutes can becompletely controlled.

Microfluidic flow controllers are traditionally externally pump-based,including hydrodynamic, reciprocating, acoustic, and peristaltic pumps,and can be as simple as a syringe (see U.S. Pat. No. 6,444,106 toMcbride et al., U.S. Pat. No. 6,811,385 to Blakley, U.S. PublishedPatent Application No. 20040028566 to Ko et al.). More recently,electroosmosis, a process that does not require moving parts, hasexperienced success as a fluid flow driver (see U.S. Pat. No. 6,406,605to Moles, U.S. Pat. No. 6,568,910 to Parse). Other fluid flow devicesthat do not require moving parts use gravity (see U.S. Pat. No.6,743,399 to Weigi et al.), centrifugal force (see U.S. Pat. No.6,632,388 to Sanae et al.), capillary action (see U.S. Pat. No.6,591,852 to McNeely et al.), or heat (see U.S. Published PatentApplication No. 20040257668 to Ito) to drive liquids through themicrochannels. Other inventions create liquid flow by the application ofan external force, such as a blade (see U.S. Pat. No. 6,068,751 toNeukermans).

Valves also are used in fluid flow control. Valves can be actuated byapplying an external force, such as a blade, cantilever, or plug to anelastomeric channel (see U.S. Pat. No. 6,068,751 to Neukermans). Elasticchannels also can contain membranes that can be deflected by airpressure and/or liquid pressure, e.g., water pressure,electrostatically, or magnetically (see U.S. Pat. No. 6,408,878 to Ungeret al.). Other 2-way valves are actuated by light (see U.S. PublishedPatent Application No. 20030156991 to Halas et al.), piezoelectriccrystals (see Published PCT International Application No. WO2003/089,138 to Davis et al.), particle deflection (see U.S. Pat. No.6,802,489 to Marr et al.), or bubbles formed within the channelelectrochemically (see Published PCT International Application No. WO2003/046,256 to Hua et al.). One-way or “check valves” also can beformed in microchannels with balls, flaps, or diaphragms (see U.S. Pat.No. 6,817,373 to Cox et al.; U.S. Pat. No. 6,554,591 to Dai et al.;Published PCT International Application No. WO 2002/053,290 to Jeon etal.). Rotary-type switching valves are used for complex reactions (seePublished PCT International Application No. WO 2002/055,188 to Powell etal.).

Microscale mixing and separation components are necessary to facilitatereactions and evaluate products. In microfluidic devices, mixing is mostoften done by diffusion, in channels of long length scales, curved, withvariable widths, or having features that cause turbulence (see U.S. Pat.No. 6,729,352 to O'Conner et al., U.S. Published Patent Application No.20030096310 to Hansen et al.). Mixing also can be accomplishedelectroosmotically (see U.S. Pat. No. 6,482,306 to Yager et al.) orultrasonically (see U.S. Pat. No. 5,639,423 to Northrup et al.).Separations in micro-scale channels typically use three methods:electrophoresis, packed columns or gel within a channel, orfunctionalization of channel walls. Electrophoresis is commonly donewith charged molecules, such as nucleic acids, peptides, proteins,enzymes, and antibodies and the like, and is the simplest technique (seeU.S. Pat. No. 5,958,202 to Regnier et al., U.S. Pat. No. 6,274,089 toChow et al.). Channel columns can be packed with porous orstationary-phase coated beads or a gel to facilitate separations (seePublished PCT International Application No. WO 2003/068,402 to Koehleret al., U.S. Published Patent Application No. 20020164816 to Quake etal., U.S. Pat. No. 6,814,859 to Koehler et al.). Possible packingmaterials include silicates, talc, Fuller's earth, glass wool, charcoal,activated charcoal, celite, silica gel, alumina, paper, cellulose,starch, magnesium silicate, calcium sulfate, silicic acid, florisil,magnesium oxide, polystyrene, p-aminobenzyl cellulose,polytetrafluoroethylene resin, polystyrene resin, SEPHADEX™ (AmershamBiosciences, Corp., Piscataway, N.J., United States of America),SEPHAROSE™ (Amersham Biosciences, Corp., Piscataway, N.J., United Statesof America), controlled pore glass beads, agarose, other solid resinsknown to one skilled in the art and combinations of two or more of anyof the foregoing. Magnetizable material, such as ferric oxide, nickeloxide, barium ferrite or ferrous oxide, also can be imbedded,encapsulated of otherwise incorporated into a solid-phase packingmaterial.

The walls of microfluidic chambers also can be functionalized with avariety of ligands that can interact or bind to an analyte or to acontaminant in an analyte solution. Such ligands include: hydrophilic orhydrophobic small molecules, steroids, hormones, fatty acids, polymers,RNA, DNA, PNA, amino acids, peptides, proteins (including antibodybinding proteins such as protein G), antibodies or antibody fragments(FABs, etc), antigens, enzymes, carbohydrates (including glycoproteinsor glycolipids), lectins, cell surface receptors (or portions thereof),species containing a positive or a negative charge, and the like (seeU.S. Published Patent Application No. 20040053237 to Liu et al.,Published PCT International Application No. WO 2004/007,582 to Augustineet al., U.S. Published Patent Application No. 20030190608 to Blackburn).

Thus, in some embodiments, the presently disclosed subject matterdescribes a method of flowing a material and/or mixing two or morematerials in a PFPE-based microfluidic device. In some embodiments, thepresently disclosed subject matter describes a method of conducting achemical reaction, including but not limited to synthesizing abiopolymer, such as DNA. In some embodiments, the presently disclosedsubject matter describes a method of screening a sample for acharacteristic. In some embodiments, the presently disclosed subjectmatter describes a method of dispensing a material. In some embodiments,the presently disclosed subject matter describes a method of separatinga material.

XI.A. Method of Flowing a Material and/or Mixing Two Materials in aPFPE-based Microfluidic Device

Referring now to FIG. 8, a schematic plan view of a microfluidic deviceof the presently disclosed subject matter is shown. The microfluidicdevice is referred to generally at 800. Microfluidic device 800 includesa patterned layer 802, and a plurality of holes 810A, 810B, 810C, and810D. These holes can be further described as inlet aperture 810A, inletaperture 810B, and inlet aperture 810C, and outlet aperture 810D. Eachof apertures 810A, 810B, 810C, and 810D are covered by seals 820A, 820B,820C, and 820D, which are preferably reversible seals. Seals 820A, 820B,820C, and 820D are provided so that materials, including but not limitedto, solvents, chemical reagents, components of a biochemical system,samples, inks, and reaction products and/or mixtures of solvents,chemical reagents, components of a biochemical system, samples, inks,reaction products and combinations thereof, can be stored, shipped, orotherwise maintained in microfluidic device 800 if desired. Seals 820A,820B, 820C, and 820D can be reversible, that is, removable, so thatmicrofluidic device 800 can be implemented in a chemical reaction orother use and then can be resealed if desired.

Continuing with reference to FIG. 8, in some embodiments, apertures810A, 810B, and 810C, further include pressure-actuated valves(including intersecting, overlaid flow channels) which can be actuatedto seal the microfluidic channel associated with the aperture.

Continuing with reference to FIG. 8, patterned layer 802 of microfluidicdevice 800 includes an integrated network 830 of microscale channels.Optionally, pattern layer 802 includes a functionalized surface, such asthat shown in FIG. 5A. Integrated network 830 can include a series offluidly connected microscale channels designated by the followingreference characters: 831, 832, 833, 834, 835, 836, 837, 838, 839, and840. Thus, inlet aperture 810A is in fluid communication with microscalechannel 831 that extends away from aperture 810A and is in fluidcommunication with microscale channel 832 via a bend. In integratednetwork 830 depicted in FIG. 8, a series of 90° bends are shown forconvenience. It is noted, however, that the paths and bends provided inthe channels of integrated network 830, can encompass any desiredconfiguration, angle, or other characteristic (such as but not limitedto a serpentine section). Indeed, fluid reservoirs 850A and 850B can beprovided along microscale channels 831, 832, 833, and 834, respectively,if desired. As shown in FIG. 8, fluid reservoirs 850A and 850B includeat least one dimension that is greater than a dimension of the channelsthat are immediately adjacent to them.

Continuing, then, with reference to FIG. 8, microscale channels 832 and834 intersect at intersecting point 860A and proceed into a singlemicroscale channel 835. Microscale channel 835 proceeds to a chamber870, which in the embodiment shown in FIG. 8, is dimensioned to be widerthan microscale channel 835. In some embodiments, chamber 870 includes areaction chamber. In some embodiments, chamber 870 includes a mixingregion. In some embodiments, chamber 870 includes a separation region.In some embodiments, the separation region includes a given dimension,e.g., length, of a channel, wherein the material is separated by charge,or mass, or combinations thereof, or any other physical characteristicwherein a separation can occur over a given dimension. In someembodiments, the separation region includes an active material 880. Aswould be understood by one of ordinary skill in the art, the term“active material” is used herein for convenience and does not imply thatthe material must be activated to be used for its intended purpose. Insome embodiments, the active material includes a chromatographicmaterial. In some embodiments, the active material includes a targetmaterial.

Continuing with FIG. 8, it is noted that chamber 870 does notnecessarily need to be of a wider dimension than an adjacent microscalechannel. Indeed chamber 870 can simply include a given segment of amicroscale channel wherein at least two materials are separated, mixed,and/or reacted. Extending from chamber 870 substantially opposite frommicroscale channel 835 is microscale channel 836. Microscale channel 836forms a T-junction with microscale channel 837, which extends away fromand is in fluid communication with aperture 810C. Thus, the junction ofmicroscale channels 836 and 837 form intersecting point 860B. Microscalechannel 838 extends from intersecting point 860B in a directionsubstantially opposite microscale channel 837 and to fluid reservoir850C. Fluid reservoir 850C is dimensioned to be wider than microscalechannel 838 for a predetermined length. As noted above, however, a givensection of a microscale channel can act as a fluid reservoir without theneed to necessarily change a dimension of the section of microscalechannel. Moreover, microscale channel 838 could act as a reactionchamber in that a reagent flowing from microscale channel 837 tointersection point 860B could react with a reagent moving frommicroscale channel 836 to intersection point 860B and into microscalechannel 838.

Continuing with reference to FIG. 8, microscale channel 839 extends fromfluid reservoir 850C substantially opposite microfluidic channel 838 andtravels through a bend into microscale channel 840. Microscale channel840 is fluidly connected to outlet aperture 810D. Outlet aperture 810Dcan optionally be reversibly sealed via seal 820D, as discussed above.Again, the reversible sealing of outlet aperture 810D can be desirablein the case of an embodiment where a reaction product is formed inmicrofluidic device 800 and is desired to be transported to anotherlocation in microfluidic device 800.

The flow of a material can be directed through the integrated network830 of microscale channels, including channels, fluid reservoirs, andreaction chambers through the use of pressure-actuated valves and thelike known in the art, for example those described in U.S. Pat. No.6,408,878 to Unger et al., which is incorporated herein by reference inits entirety. The presently disclosed subject matter thus provides amethod of flowing a material through a PFPE-based microfluidic device.In some embodiments, the method includes providing a microfluidic deviceincluding (i) a perfluoropolyether (PFPE) material having acharacteristic selected from the group consisting of: a viscositygreater than about 100 centistokes (cSt); a viscosity less than about100 cSt, provided that the liquid PFPE precursor material having aviscosity less than 100 cSt is not a free-radically photocurable PFPEmaterial; (ii) a functionalized PFPE material; (iii) afluoroolefin-based elastomer; and (iv) combinations thereof, and whereinthe microfluidic device includes one or more microscale channels; andflowing a material in the microscale channel.

Also provided is a method of mixing two or more materials. In someembodiments, the method includes providing a microscale device including(i) a perfluoropolyether (PFPE) material having a characteristicselected from the group consisting of: a viscosity greater than about100 centistokes (cSt); a viscosity less than about 100 cSt, providedthat the liquid PFPE precursor material having a viscosity less than 100cSt is not a free-radically photocurable PFPE material; (ii) afunctionalized PFPE material; (iii) a fluoroolefin-based elastomer; and(iv) combinations thereof; and contacting a first material and a secondmaterial in the device to mix the first and second materials.Optionally, the microscale device is selected from the group consistingof a microfluidics device and a microtiter plate.

In some embodiments, the method includes disposing a material in themicrofluidic device. In some embodiments, as is best shown in FIG. 10and as discussed in more detail herein below, the method includesapplying a driving force to move the material along the microscalechannel.

In some embodiments, the layer of PFPE material covers a surface of atleast one of the one or more microscale channels. Optionally, the layerof PFPE material includes a functionalized surface. In some embodiments,the microfluidic device includes one or more patterned layers of PFPEmaterial, and wherein the one or more patterned layers of the PFPEmaterial defines the one or more microscale channels. In this case thepatterned layer of PFPE can include a functionalized surface. In someembodiments, the microfluidic device can further include a patternedlayer of a second polymeric material, wherein the patterned layer of thesecond polymeric material is in operative communication with the atleast one of the one or more patterned layers of PFPE material. See FIG.2.

In some embodiments, the method includes at least one valve. In someembodiments the valve is a pressure-actuated valve, wherein thepressure-actuated valve is defined by one of: (a) a microscale channel;and (b) at least one of the plurality of holes. In some embodiments, thepressure-actuated valve is actuated by introducing a pressurized fluidor a gas that does not permeate the material including the microscalechannel (e.g., Sulfur Hexafluoride) into one of: (a) a microscalechannel; and (b) at least one of the plurality of holes.

In some embodiments, the pressurized fluid has a pressure between about10 psi and about 60 psi. In some embodiments, the pressure is about 25psi. In some embodiments, the material includes a fluid. In someembodiments, the fluid includes a solvent. In some embodiments, thesolvent includes an organic solvent. In some embodiments, the materialflows in a predetermined direction along the microscale channel.

In the case of mixing two materials, which in some embodiments caninclude mixing two reactants to provide a chemical reaction, thecontacting of the first material and the second material is performed ina mixing region defined in the one or more microscale channels. Themixing region can include a geometry selected from the group consistingof a T-junction, a serpentine, an elongated channel, a microscalechamber, and a constriction. Optionally, the first material and thesecond material are disposed in separate channels of the microfluidicdevice. Also, the contacting of the first material and the secondmaterial can be performed in a mixing region defined by an intersectionof the channels.

Continuing with a method of mixing, the method can include flowing thefirst material and the second material in a predetermined direction inthe microfluidic device, and can include flowing the mixed materials ina predetermined direction in the microfluidic device. In someembodiments, the mixed material can be contacted with a third materialto form a second mixed material. In some embodiments the mixed materialincludes a reaction product and the reaction product can be subsequentlyreacted with a third reagent. One of ordinary skill in the art uponreview of the presently disclosed subject matter would recognize thatthe description of the method of mixing provided immediately hereinaboveis for the purposes of illustration and not limitation. Accordingly, thepresently disclosed method of mixing materials can be used to mix aplurality of materials and form a plurality of mixed materials and/or aplurality of reaction products. The mixed materials, including but notlimited to reaction products, can be flowed to an outlet aperture of themicrofluidic device. A driving force can be applied to move thematerials through the microfluidic device. See FIG. 10. In someembodiments the mixed materials are recovered.

In an embodiment employing a microtiter plate, the microtiter plate caninclude one or more wells. In some embodiments, the layer of PFPEmaterial covers a surface of at least one of the one or more wells. Thelayer of PFPE material can include a functionalized surface. See FIG.5B.

XI.B. Method of Synthesizing a Biopolymer in a PFPE-based MicrofluidicDevice

In some embodiments, the presently disclosed PFPE-based microfluidicdevice can be used in biopolymer synthesis, for example, in synthesizingoligonucleotides, proteins, peptides, DNA, and the like. In someembodiments, such biopolymer synthesis systems include an integratedsystem including an array of reservoirs, fluidic logic for selectingflow from a particular reservoir, an array of channels, reservoirs, andreaction chambers in which synthesis is performed, and fluidic logic fordetermining into which channels the selected reagent flows.

Referring now to FIG. 9, a plurality of reservoirs, e.g., reservoirs910A, 910B, 910C, and 910D, have bases A, C, T, and G respectivelydisposed therein, as shown. Four flow channels 920A, 920B, 920C, and920D are connected to reservoirs 910A, 910B, 910C, and 910D. Fourcontrol channels 922A, 922B, 922C, and 922D (shown in phantom) aredisposed thereacross with control channel 922A permitting flow onlythrough flow channel 920A (i.e., sealing flow channels 920B, 920C, and920D), when control channel 922A is pressurized. Similarly, controlchannel 922B permits flow only through flow channel 920B whenpressurized. As such, the selective pressurization of control channels922A, 922B, 922C, and 922D sequentially selects a desired base A, C, T,and G from a desired reservoir 910A, 910B, 910C, or 910D. The fluid thenpasses through flow channel 920E into a multiplexed channel flowcontroller 930, (including, for example, any system as shown in FIG. 8)which in turn directs fluid flow into one or more of a plurality ofsynthesis channels or reaction chambers 940A, 940B, 940C, 940D, or 940Ein which solid phase synthesis can be carried out.

In some embodiments, instead of starting from the desired base A, C, T,and G, a reagent selected from one of a nucleotide and a polynucleotideis disposed in at least one of reservoir 910A, 910B, 910C, and 910D. Insome embodiments, the reaction product includes a polynucleotide. Insome embodiments, the polynucleotide is DNA.

Accordingly, after a review of the present disclosure, one of ordinaryskill in the art would recognize that the presently disclosed PFPE-basedmicrofluidic device can be used to synthesize biopolymers, as describedin U.S. Pat. No. 6,408,878 to Unger et al. and U.S. Pat. No. 6,729,352to O'Conner et al., and/or in a combinatorial synthesis system asdescribed in U.S. Pat. No. 6,508,988 to van Dam et al., each of which isincorporated herein by reference in its entirety.

XI.C. Method of Incorporating a PFPE-Based Microfluidic Device into anIntegrated Fluid Flow System

In some embodiments, the method of performing a chemical reaction orflowing a material within a PFPE-based microfluidic device includesincorporating the microfluidic device into an integrated fluid flowsystem. Referring now to FIG. 10, a system for carrying out a method offlowing a material in a microfluidic device and/or a method ofperforming a chemical reaction in accordance with the presentlydisclosed subject matter is schematically depicted. The system itself isgenerally referred to at 1000. System 1000 can include a centralprocessing unit 1002, one or more driving force actuators 1010A, 1010B,1010C, and 1010D, a collector 1020, and a detector 1030. In someembodiments, detector 1030 is in fluid communication with themicrofluidic device (shown in shadow). System microfluidic device 1000of FIG. 8, and these reference numerals of FIG. 8 are employed in FIG.10. Central processing unit (CPU) 1002 can be, for example, a generalpurpose personal computer with a related monitor, keyboard or otherdesired user interface. Driving force actuators 1010A, 1010B, 1010C, and1010D can be any suitable driving force actuator as would be apparent toone of ordinary skill in the art upon review of the presently disclosedsubject matter. For example, driving force actuators 1010A, 1010B,1010C, and 1010D can be pumps, electrodes, injectors, syringes, or othersuch devices that can be used to force a material through a microfluidicdevice. Representative driving forces themselves thus include capillaryaction, pump driven fluid flow, electrophoresis based fluid flow, pHgradient driven fluid flow, or other gradient driven fluid flow.

In the schematic of FIG. 10 driving force actuator 1010D is shown asconnected at outlet aperture 810D, as will be described below, todemonstrate that at least a portion of the driving force can be providedat the end point of the desired flow of solution, reagent, and the like.Collector 1020 also is provided to show that a reaction product 1048, asdiscussed below, can be collected at the end point of system flow. Insome embodiments, collector 1020 includes a fluid reservoir. In someembodiments, collector 1020 includes a substrate. In some embodiments,collector 1020 includes a detector. In some embodiments, collector 1020includes a subject in need of therapeutic treatment. For convenience,system flow is generally represented in FIG. 10 by directional arrowsF1, F2, and F3.

Continuing with reference to FIG. 10, in some embodiments a chemicalreaction is performed in integrated flow system 1000. In someembodiments, material 1040, e.g., a chemical reagent, is introduced tomicrofluidic device 1000 through aperture 810A, while a second material1042, e.g., a second chemical reagent, is introduced to microfluidicdevice 1000, via inlet aperture 810B. Optionally, microfluidics device1000 includes a functionalized surface (see FIG. 5A). Driving forceactuators 1010A and 1010B propel chemical reagents 1040 and 1042 tomicrofluidic channels 831 and 833, respectively. Flow of chemicalreagents 1040 and 1042 continues to fluid reservoirs 850A and 850B,where a reserve of reagents 1040 and 1042 is collected. Flow of chemicalreagents 1040 and 1042 continues into microfluidic channels 832 and 834to intersection point 860A wherein initial contact between chemicalreagents 1040 and 1042 occurs. Flow of chemical reagents 1040 and 1042then continues to reaction chamber 870 where a chemical reaction betweenchemical reagents 1040 and 1042 proceeds.

Continuing with reference to FIG. 10, reaction product 1044 flows tomicroscale channel 836 and to intersection point 860B. Chemical reagent1046 then reacts with reaction product 1044 beginning at intersectionpoint 860B through reaction chamber 838 and to fluid reservoir 850C. Asecond reaction product 1048 is formed. Flow of the second reactionproduct 1048 continues through microscale channel 840 to aperture 810Dand finally into collector 1020. Thus, it is noted that CPU 1002actuates driving force actuator 1010C such that chemical reagent 1046 isreleased at an appropriate time to contact reaction product 1044 atintersection point 860B.

XI.D. Representative Applications of a Microfluidic Device

In some embodiments, the presently disclosed subject matter discloses amethod of screening a sample for a characteristic. In some embodiments,the presently disclosed subject matter discloses a method of dispensinga material. In some embodiments, the presently disclosed subject matterdiscloses a method of separating a material. Accordingly, one ofordinary skill in the art would recognize that a microfluidic devicedescribed herein can be applied to many applications, including, but notlimited to, genome mapping, rapid separations, sensors, nanoscalereactions, ink-jet printing, drug delivery, Lab-on-a-Chip, in vitrodiagnostics, injection nozzles, biological studies, high-throughputscreening technologies, such as for use in drug discovery and materialsscience, diagnostic and therapeutic tools, research tools, and thebiochemical monitoring of food and natural resources, such as soil,water, and/or air samples collected with portable or stationarymonitoring equipment. In some embodiments, the presently disclosedsubject matter discloses screening a material through a microfluidicdevice. Some useful microfluidic devices suitable for ultravioletdetection are described in International Patent Publication Number WO02/29397 A2, which is incorporated herein by reference in its entirety.

XI.D.1. Method of Screening a Sample for a Characteristic

In some embodiments, the presently disclosed subject matter discloses amethod of screening a sample for a characteristic. In some embodiments,the method includes:

(a) providing a microscale device comprising:

-   -   (i) a perfluoropolyether (PFPE) material having a characteristic        selected from the group consisting of: a viscosity greater than        about 100 centistokes (cSt) and a viscosity less than about 100        cSt, provided that the liquid PFPE precursor material having a        viscosity less than 100 cSt is not a free-radically photocurable        PFPE material;    -   (ii) a functionalized PFPE material;    -   (iii) a fluoroolefin-based elastomer; and    -   (iv) combinations thereof;

(b) providing a target material;

(c) disposing the sample in the microscale device;

(d) contacting the sample with the target material; and

(e) detecting an interaction between the sample and the target,

wherein the presence or the absence of the interaction is indicative ofthe characteristic of the sample.

Referring once again to FIG. 10, at least one of materials 1040 and 1042includes a sample. In some embodiments, at least one of materials 1040and 1042 includes a target material. Thus, a “sample” generally refersto any material about which information relating to a characteristic isdesired. Also, a “target material” can refer to any material that can beused to provide information relating to a characteristic of a samplebased on an interaction between the target material and the sample. Insome embodiments, for example, when sample 1040 contacts target material1042 an interaction occurs. In some embodiments, the interactionproduces a reaction product 1044. In some embodiments, the interactionincludes a binding event. In some embodiments, the binding eventincludes the interaction between, for example, an antibody and anantigen, an enzyme and a substrate, or more particularly, a receptor anda ligand, or a catalyst and one or more chemical reagents. In someembodiments, the reaction product is detected by detector 1030.

In some embodiments, the method includes disposing the target materialin at least one of the plurality of channels. Referring once again toFIG. 10, in some embodiments, the target material includes activematerial 880. In some embodiments, the target material, the sample, orboth the target and the sample are bound to a functionalized surface. Insome embodiments, the target material includes a substrate, for examplea non-patterned layer. In some embodiments, the substrate includes asemiconductor material. In some embodiments, at least one of theplurality of channels of the microfluidic device is in fluidcommunication with the substrate, e.g., a non-patterned layer. In someembodiments, the target material is disposed on a substrate, e.g., anon-patterned layer. In some embodiments, at least one of the one ormore channels of the microfluidic device is in fluid communication withthe target material disposed on the substrate.

In some embodiments, the method includes disposing a plurality ofsamples in at least one of the plurality of channels. In someembodiments, the sample is selected from the group consisting of atherapeutic agent, a diagnostic agent, a research reagent, a catalyst, ametal ligand, a non-biological organic material, an inorganic material,a foodstuff, soil, water, and air. In some embodiments, the sampleincludes one or more members of one or more libraries of chemical orbiological compounds or components. In some embodiments, the sampleincludes one or more of a nucleic acid template, a sequencing reagent, aprimer, a primer extension product, a restriction enzyme, a PCR reagent,a PCR reaction product, or a combination thereof. In some embodiments,the sample includes one or more of an antibody, a cell receptor, anantigen, a receptor ligand, an enzyme, a substrate, an immunochemical,an immunoglobulin, a virus, a virus binding component, a protein, acellular factor, a growth factor, an inhibitor, or a combinationthereof.

In some embodiments, the target material includes one or more of anantigen, an antibody, an enzyme, a restriction enzyme, a dye, afluorescent dye, a sequencing reagent, a PCR reagent, a primer, areceptor, a ligand, a chemical reagent, or a combination thereof.

In some embodiments, the interaction includes a binding event. In someembodiments, the detecting of the interaction is performed by at leastone or more of a spectrophotometer, a fluorometer, a photodiode, aphotomultiplier tube, a microscope, a scintillation counter, a camera, aCCD camera, film, an optical detection system, a temperature sensor, aconductivity meter, a potentiometer, an amperometric meter, a pH meter,or a combination thereof.

Accordingly, after a review of the present disclosure, one of ordinaryskill in the art would recognize that the presently disclosed PFPE-basedmicrofluidic device can be used in various screening techniques, such asthose described in U.S. Pat. No. 6,749,814 to Bergh et al., U.S. Pat.No. 6,737,026 to Bergh et al., U.S. Pat. No. 6,630,353 to Parce et al.,U.S. Pat. No. 6,620,625 to Wolk et al., U.S. Pat. No. 6,558,944 to Parceet al., U.S. Pat. No. 6,547,941 to Kopf-Sill et al., U.S. Pat. No.6,529,835 to Wada et al., U.S. Pat. No. 6,495,369 to Kercso et al., andU.S. Pat. No. 6,150,180 to Parce et al., each of which is incorporatedby reference in its entirety. Further, after a review of the presentdisclosure, one of ordinary skill in the art would recognize that thepresently disclosed PFPE-based microfluidic device can be used, forexample, to detect DNA, proteins, or other molecules associated with aparticular biochemical system, as described in U.S. Pat. No. 6,767,706to Quake et al., which is incorporated herein by reference in itsentirety.

XI.D.2. Method of Dispensing a Material

Additionally, the presently disclosed subject matter describes a methodof dispensing a material. In some embodiments, the method includes:

(a) providing a microfluidic device comprising:

-   -   (i) a perfluoropolyether (PFPE) material having a characteristic        selected from the group consisting of: a viscosity greater than        about 100 centistokes (cSt) and a viscosity less than about 100        cSt, provided that the liquid PFPE precursor material having a        viscosity less than 100 cSt is not a free-radically photocurable        PFPE material;    -   (ii) a functionalized PFPE material;    -   (iii) a fluoroolefin-based elastomer; and    -   (iv) combinations thereof; and wherein the microfluidics device        includes one or more microscale channels, and wherein at least        one of the one or more microscale channels includes an outlet        aperture;

(b) providing at least one material;

(c) disposing at least one material in at least one of the one or moremicroscale channels; and

(d) dispensing at least one material through the outlet aperture.

In some embodiments, the layer of PFPE material covers a surface of atleast one of the one or more microscale channels.

Referring once again to FIG. 10, in some embodiments, a material, e.g.,material 1040, second material 1042, chemical reagent 1046, reactionproduct 1044, and/or reaction product 1048 flow through outlet aperture810D and are dispensed in or on collector 1020. In some embodiments, thetarget material, the sample, or both the target and the sample are boundto a functionalized surface.

In some embodiments, the material includes a drug. In some embodiments,the method includes metering a predetermined dosage of the drug. In someembodiments, the method includes dispensing the predetermined dosage ofthe drug.

In some embodiments, the material includes an ink composition. In someembodiments, the method includes dispensing the ink composition on asubstrate. In some embodiments, the dispensing of the ink composition ona substrate forms a printed image.

Accordingly, after a review of the present disclosure, one of ordinaryskill in the art would recognize that the presently disclosed PFPE-basedmicrofluidic device can be used for microfluidic printing as describedin U.S. Pat. No. 6,334,676 to Kaszczuk et al., U.S. Pat. No. 6,128,022to DeBoer et al., and U.S. Pat. No. 6,091,433 to Wen, each of which isincorporated herein by reference in its entirety.

XI.D.3 Method of Separating a Material

In some embodiments, the presently disclosed subject matter describes amethod of separating a material, the method includes:

(a) providing a microfluidic device comprising:

-   -   (i) a perfluoropolyether (PFPE) material having a characteristic        selected from the group consisting of: a viscosity greater than        about 100 centistokes (cSt) and a viscosity less than about 100        cSt, provided that the liquid PFPE precursor material having a        viscosity less than 100 cSt is not a free-radically photocurable        PFPE material;    -   (ii) a functionalized PFPE material;    -   (iii) a fluoroolefin-based elastomer; and    -   (iv) combinations thereof; and wherein the microfluidics device        includes one or more microscale channels, and wherein at least        one of the one or more microscale channels includes a separation        region;

(b) disposing a mixture comprising at least a first material and asecond material in the microfluidic device;

(c) flowing the mixture through the separation region; and

(d) separating the first material from the second material in theseparation region to form at least one separated material.

Referring once again to FIG. 10, in some embodiments, at least one ofmaterial 1040 and second material 1042 include a mixture. For example,material 1040, e.g., a mixture, flows through the microfluidic system tochamber 870, which in some embodiments includes a separation region. Insome embodiments, the separation region includes active material 880,e.g., a chromatographic material. Material 1040, e.g., a mixture, isseparated in chamber 870, e.g., a separation chamber, to form a thirdmaterial 1044, e.g., a separated material. In some embodiments,separated material 1044 is detected by detector 1030.

In some embodiments, the separation region includes a chromatographicmaterial. In some embodiments, the chromatographic material is selectedfrom the group consisting of a size-separation matrix, anaffinity-separation matrix, and a gel-exclusion matrix, or a combinationthereof.

In some embodiments, the first or second material includes one or moremembers of one or more libraries of chemical or biological compounds orcomponents. In some embodiments, the first or second material includesone or more of a nucleic acid template, a sequencing reagent, a primer,a primer extension product, a restriction enzyme, a PCR reagent, a PCRreaction product, or a combination thereof. In some embodiments, thefirst or second material includes one or more of an antibody, a cellreceptor, an antigen, a receptor ligand, an enzyme, a substrate, animmunochemical, an immunoglobulin, a virus, a virus binding component, aprotein, a cellular factor, a growth factor, an inhibitor, or acombination thereof.

In some embodiments, the method includes detecting the separatedmaterial. In some embodiments, the detecting of the separated materialis performed by at least one or more of a spectrophotometer, afluorometer, a photodiode, a photomultiplier tube, a microscope, ascintillation counter, a camera, a CCD camera, film, an opticaldetection system, a temperature sensor, a conductivity meter, apotentiometer, an amperometric meter, a pH meter, or a combinationthereof.

Accordingly, after a review of the present disclosure, one of ordinaryskill in the art would recognize that the presently disclosed PFPE-basedmicrofluidic device can be used to separate materials, as described inU.S. Pat. No. 6,752,922 to Huang et al., U.S. Pat. No. 6,274,089 to Chowet al., and U.S. Pat. No. 6,444,461 to Knapp et al., each of which isincorporated herein by reference in its entirety.

XII. Applications for Functionalized Microfluidic Devices

Fluidic microchip technologies are increasingly being used asreplacements for traditional chemical and biological laboratoryfunctions. Microchips that perform complex chemical reactions,separations, and detection on a single device have been fabricated.These “lab-on-a-chip” applications facilitate fluid and analytetransport with the advantages of reduced time and chemical consumptionand ease of automation.

A variety of biochemical analysis, reactions, and separations have beenperformed within microchannel systems. High throughput screening assaysof synthesized molecules and natural products are of great interest.Microfluidic devices for screening a wide variety of molecules based ontheir ability to inhibit the interactions of enzymes and fluorescentlylabeled substrates have been described (U.S. Pat. No. 6,046,056, toParse et al.). As described by Parse et al., such devices allow forscreening natural or synthetic libraries of potential drugs throughtheir antagonist or agonist properties. The types of molecules that canbe screened include, but are not limited to, small organic or inorganicmolecules, polysaccharides, peptides, proteins, nucleic acids orextracts of biological materials such as bacteria, fungi, yeast, plantsand animal cells. The analyte compounds can be free in solution orattached to a solid support, such as agarose, cellulose, dextran,polystyrene, carboxymethyl cellulose, polyethylene glycol (PEG), filterpaper, nitrocellulose, ion exchange resins, plastic films, glass beads,polyaminemethylvinylether maleic acid copolymer, amino acid copolymer,ethylene-maleic acid copolymer, nylon, silk, and the like. Compounds canbe tested as pure compounds or in pools. For example, U.S. Pat. No.6,007,690 to Nelson et al. relates to a microfluidic moleculardiagnostic that purifies DNA from whole blood samples. The device usesan enrichment channel that cleans up or concentrates the analyte sample.For example, the enrichment channel could hold antibody coated beads toremove various cell parts via their antigenic components or could holdchromatographic components, such as ion exchange resin or a hydrophobicor hydrophilic membrane. The device also can include a reactor chamber,wherein various reactions can be performed on the analyte, such as alabeling reaction or in the case of a protein analyte, a digestionreaction. Further, U.S. Published Patent Application No. 20040256570 toBeebe et al. describes a device where antibody interaction with anantigenic analyte material coated on the outside of a liposome isdetected when that interaction causes the lysis of the liposome and itsrelease of a detectable molecule. U.S. Published Patent Application No.20040132166 to Miller et al. provides a microfluidic device that cansense environmental factors, such as pH, humidity, and O₂ levelscritical for the growth of cells. The reaction chambers in these devicescan function as bioreactors capable of growing cells, allowing for theiruse to transfect cells with DNA and produce proteins, or to test for thepossible bioavailability of drug substances by measuring theirabsorbance across CACO-2 cell layers.

In addition of growing cells, microfluidic devices also have been usedto sort cells. U.S. Pat. No. 6,592,821 to Wada et al. describeshydrodynamic focusing to sort cells and subcellular components,including individual molecules, such as nucleic acids, polypeptides orother organic molecules, or larger cell components like organelles. Themethod can sort for cell viability or other cellular expressionfunctions.

Amplification, separation, sequencing, and identification of nucleicacids and proteins are common microfluidic device applications. Forexample, U.S. Pat. No. 5,939,291 to Loewy et al. illustrate amicrofluidic device that uses electrostatic techniques to performisothermal nucleic acid amplification. The device can be used inconjunction with a number of common amplification reaction strategies,including PCR (polymerase chain reaction), LCR (ligase chain reaction),SDA (strand displacement amplification), NASBA (nucleic acidsequence-based amplification), and TMA (transcription-mediatedamplification). U.S. Pat. No. 5,993,611 to Moroney et al. describes adevice that uses capacitive charging to analyze, amplify or otherwisemanipulate nucleic acids. Devices have been designed that sort DNA bysize, analyzing restriction fragment length polymorphism (see U.S. Pat.No. 6,833,242 to Quake et al.). The devices also can have particular usein forensic applications, such as DNA fingerprinting. U.S. Pat. No.6,447,724 to Jensen et al. describes microfluidics that identifycomponents of a mixture based on the different fluorescent lifetimes ofthe labels attached to members of the mixture. Such a device could beused to analyze sequencing reactions of nucleic acids, proteins oroligosaccharides or to inspect or interrogate members of a combinatoriallibrary of organic molecules.

Other microfluidic devices directed toward specific protein applicationsinclude a device that promotes protein crystal growth in microfluidicchannels (see U.S. Pat. No. 6,409,832, to Weigl et al.). In the device,protein sample and solvent are directed to a channel with laminar flowcharacteristics that form diffusion zones, which provide well-definedcrystallization. U.S. Published Patent Application No. 2004/0121449 toPugia et al. illustrates a device that can separate red blood cells fromplasma using minimal centrifugal force on sample sizes as small as 5microliters. The device could be particularly useful in clinicaldiagnostics and also could be used to separate any particulate matterfrom a liquid.

As partly described hereinabove, microfluidic devices have been utilizedas microreactors for a variety of chemical and biological applications.Chambers in these devices can be used for sequencing, restriction enzymedigests, restriction fragment length polymorphism (RFLP) analysis,nucleic acid amplification, or gel electrophoresis (see U.S. Pat. No.6,130,098, to Handigue et al.). A multitude of chemical titrationreactions can be run in the devices (see U.S. Published PatentApplication No. 20040258571, to Lee et al.), including acid-basedtitrations or titrations based on precipitation (for example, Ag(I) withCl⁻, Br⁻, I⁻, or SCN⁻), complex formation (for example, Ag(I) with CN⁻),or redox reactions (such as Fe(II)/Fe(III) with Ce(III)/Ce(IV)).Further, a sensor for potentiometry, amperometry, spectrophotometry,turbidometry, fluorimetry or calorimetry can be attached to the device.Fractionation of proteins (see U.S. Published Patent Application No.20040245102, to Gilbert et al.) based physical or biological propertiesis of use in protein expression analysis (finding molecular markers,determining a molecular basis or profile for a disease state orinterpreting protein structure/function relationships). A variety ofelectrophoresis techniques (including capillary isoelectric focusing,capillary zone electrophoresis, and capillary gel electrophoresis) havebeen employed in microfluidic devices for fractionating proteins (seeU.S. Pat. No. 6,818,112, to Schneider et al.). The differentelectrophoretic techniques can be used in series, with or without alabeling step to help with quantitation, and in conjunction with avariety of elution techniques (such as hydrodynamic salt mobilization,pH mobilization, or electroosmotic flow) to further separate proteins. Avariety of other materials have been used to aid in separation processesin microfluidic devices. Such materials can be attached to channel wallsin a device or be present as a separate matrix inside a channel (seeU.S. Pat. No. 6,581,441 to Paul; U.S. Pat. No. 6,613,581, to Wada etal.). Parallel separation channels can exist to separate many samples atthe same time. The solid separation media can be present as a discreteparticle or as a porous monolithic solid. Possible materials includesilica gel, agarose-based gels, polyacrylamide gels, a colloidalsolution, such as a gelatin, starches, non-ionic macroreticular andmacroporous resins (such as AMBERCHROM™ (Rohm and Haas Co, Philadelphia,Pa., United States of America), AMBERLITE™ (Rohm and Haas Co,Philadelphia, Pa., United States of America), DOWEX™ (The Dow ChemicalCompany, Midland, Mich., United States of America), DUOLITE® (Rohm andHaas Co, Philadelphia, Pa., United States of America), and the like), ormaterial present as beads (glass, metal, silica, acrylic, SEPHAROSE™,cellulose, ceramic, polymer, and the like). These materials also canhave present on their surfaces various biologically based molecules toaid in separation (for example, lectins bind to carbohydrates andantibodies can bind to antigenic groups on different proteins).Membranes within microchannels have been used for electroosmoticseparation (see U.S. Pat. No. 6,406,605, to Moles). Suitable membranescan include materials, such as track etched polycarbonate or polyimide.

Temperature, concentration and flow gradients also have been employed toaid in separation in microfluidic devices. U.S. Published PatentApplication No. 20040142411 to Kirk et al. discloses the use ofchemotaxis (the movement of cells induced by a concentration gradient ofa soluble chemotactic stimulus), hapatotaxis (the movement of cells inresponse to a concentration gradient of a substrate-bound stimulus) andchemoinvasion (the movement of cells into and/or through a barrier orgel matrix in response to a stimulus). Chemotatic stimuli includechemorepellants and chemoattractants. A chemoattractant is any substancethat attracts cells. Examples include, but are not limited to, hormonessuch as epinephrine and vasopressin; immunological agents such asinterleukein-2; growth factors, chemokines, cytokines, and variouspeptides, small molecules and cells. Chemorepellants include irritantssuch as benzalkonium chloride, propylene glycol, methanol, acetone,sodium dodecyl sulfate, hydrogen peroxide, 1-butanol, ethanol anddimethylsulfoxide; toxins, such as cyanide, carbonylcyanidechlorophenylhydrozone; endotoxins and bacterial lipopolysaccharides;viruses; pathogens; and pyrogens. Non-limiting examples of cells thatcan be manipulated by these techniques include lymphocytes, monocytes,leukocytes, macrophages, mast cells, T-cells, B-cells, neutrophils,basophils, fibroblasts, tumor cells and many others.

Microfluidic devices as sensors have garnered attention in the last fewyears. Such microfluidic sensors can include dye-based detectionsystems, affinity-based detections systems, microfabricated gravimetricanalyzers, CCD cameras, optical, detectors, optical microscopy systems,electrical systems, thermocouples, thermoresistors, and pressuresensors. Such devices have been used to detect biomolecules (seePublished PCT International Application No. WO 2004/094,986 to Althauset al.), including polynucleotides, proteins and viruses through theirinteraction with probe molecules capable of providing an electrochemicalsignal. For example, intercalation of a nucleic acid sample with a probemolecule, such as doxorubicin can reduce the amount of free doxorubicinin contact with an electrode; and a change in electrical signal results.Devices have been described that contain sensors for detecting andcontrolling environmental factors inside device reaction chambers suchas humidity, pH, dissolved O₂ and dissolved CO₂ (see Published PCTInternational Application No. WO 2004/069,983 to Rodgers et al.). Suchdevices have particular use in growing and maintaining cells. The carboncontent of samples can be measured in a device (see U.S. Pat. No.6,444,474 to Thomas et al.) wherein UV irradiation oxidizes organics toCO₂, which is then quantitated by conductivity measurements or infraredmethods. Capacitance sensors used in microfluidic devices (see PublishedPCT International Application No. WO 2004/085,063 to Xie et al.) can beused to measure pressure, flow, fluid levels, and ion concentrations.

Another application for microfluidic systems includes the highthroughput injection of cells (see Published PCT InternationalApplication No. WO 00/20554 to Garman et al.) In such a device, cellsare impelled to a needle where they can be injected with a wide varietyof materials including molecules and macromolecules, genes, chromosomes,or organelles. The device also can be used to extract material fromcells and would be of use in a variety of fields, such as gene therapy,pharmaceutical or agrochemical research, and diagnostics. Microfluidicdevices also have been used as a means of delivering ink in ink-jetprinting (see U.S. Pat. No. 6,575,562 to Anderson et al.), and to directsample solutions onto an electrospray ionization tip for massspectrometry (see U.S. Pat. No. 6,803,568 to Bousse et al.). Systems fortransdermal drug delivery also have been reported (see Published PCTInternational Application No. WO 2002/094,368 to Cormier et al.), aswell as devices containing light altering elements for use inspectroscopy applications (see U.S. Pat. No. 6,498,353 to Nagle et al.).

XIII. Applications for Functionalized Microtiter Plates

The presently disclosed materials and methods also can be applied to thedesign and manufacture of devices to be used in the manner of microtiterplates. Microtiter plates have a variety of uses in the fields of highthroughput screening for proteomics, genomics and drug discovery,environmental chemistry assays, parallel synthesis, cell culture,molecular biology and immunoassays. Common base materials used formicrotiter plates include hydrophobic materials, such as polystyrene andpolypropylene, and hydrophilic materials, such as glass. Silicon, metal,polyester, polyolefin and polytetrafluoroethylene surfaces also havebeen used for microtiter plates.

Surfaces can be selected for a particular application based on theirsolvent and temperature compatibilities and for their ability (or lackof ability) to interact with the molecules or biomolecules being assayedor otherwise manipulated. Chemical modification of the base material isoften useful in tailoring the microtiter plate to its desired functioneither by modifying the surface characteristics or by providing a sitefor the covalent attachment of a molecule or biomolecule. Thefunctionalizable nature of the presently disclosed materials is wellsuited for these purposes.

Some applications call for surfaces with low binding characteristics.Proteins and many other biomolecules (such as eukaryotic and microbialcells) can passively adsorb to polystyrene through hydrophobic or ionicinteractions. Some surface-modified base materials have been developedto address this problem. CORNING® Ultra Low Attachment (CorningIncorporated—Life Sciences, Acton, Mass., United States of America) is ahydrogel-coated polystyrene. The hydrogel coating renders the surfaceneutral and hydrophilic, preventing the attachment of almost all cells.Vessels made from the surface have uses in preventing serum proteinabsorption, in preventing anchorage-dependent cells (MDCK, VERO, C6, andthe like) from dividing, in selectively culturing tumor or virallytransformed cells as unattached colonies, in preventing stem cells fromattachment-mediated differentiation, and in studying the activation andinactivation mechanisms of macrophages. NUNC MINISORP™ (Nalgene NuncInternational, Naperville, Ill., United States of America) ispolyethylene-based product with low protein affinity and has uses forDNA probe and serum-based assays where non-specific binding is aproblem.

For other applications base, materials have been modified to enhancetheir ability to adhere to cells and other biomolecules. NUNCLON Δ™(Nalgene Nunc International) is a polystyrene surface treated by coronaor plasma discharge to add surface carboxyl groups, rendering thematerial hydrophilic and negatively charged. The material has been usedin the cell culture of a variety of cells. Polyolefin and polyestermaterials also have been treated to enhance their hydrophilicity andthereby become good surfaces for the adhesion and growth of cells (forexample PERMANOX™ and THERMANOX™, also from Nalgene Nunc International).Base materials can be coated with poly-D-lysine, collagen or fibronectinto create a positively charged surface, which also can enhance cellattachment, growth and differentiation.

Further, other molecules can be absorbed to a microtiter-like plate.Nunc MAXISORP™ (Nalgene Nunc) is a modified polystyrene base that has ahigh affinity for polar molecules and is recommended for surfaces whereantibodies need to be absorbed to the surface, as in the case of manyELISA assays. Surfaces also can be modified to interact with analytes ina more specific manner. Examples of such functional modificationsinclude nickel-chelate modified surfaces for the quantification anddetection of histidine-tagged fusion proteins and glutathione-modifiedsurfaces for the capture of GST-tagged fusion proteins.Streptavidin-coated surfaces can be used when working with biotinylatedproteins.

Some modified surfaces provide sites for the covalent attachment ofvarious molecules or biomolecules. COVALINK™ NH Secondary Amine surface(Nalgene Nunc International) is a polystyrene surface covered withsecondary amines which can bind proteins and peptides through theircarboxyl groups via carbodimide chemistry or bind DNA through theformation of a 5′ phosphoramidiate bond (again using carbodimidechemistry). Other molecules, carbohydrates, hormones, small moleculesand the like, containing or modified to contain carboxylate groups alsocan be bound to the surface. Epoxide is another useful moiety forcovalently linking groups to surfaces. Epoxide modified surfaces havebeen used to create DNA chips via the reaction of amino-modifiedoligonucleotides with surfaces. Surfaces with immobilizedoligonucleotides can be of use in high throughput DNA and RNA detectionsystems and in automated DNA amplification applications.

Other uses for microtiter plates are directed toward modifying thesurface to make it more hydrophobic, rendering it more compatible withorganic solvents or to reduce the absorption of drugs, usually smallorganic molecules. For example, Total Drug Analysis assays generallyrely on using acetonitrile to precipitate proteins and salts from aplasma or serum sample. The drug being assayed must remain in solutionfor subsequent quantification. Organic solvent-compatible microtiterplates also have uses as high performance liquid chromatography (HPLC)or liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS)prep devices and as combinatorial chemistry or parallel synthesisreaction vessels (either for solution-based or solid phase chemistries).Examples of surfaces for these types of uses include MULTICHEM™microplates (Whatman, Inc., Florham Park, N.J., United States ofAmerica) and MULTISCREEN® Solvinert (Millipore, Billerica, Mass., UnitedStates of America).

XIV. Method for Using a Functionalized Perfluoropolyether Network as aGas Separation Membrane

The presently disclosed subject matter provides for the use of afunctionalized perfluoropolyether (PFPE) network as a gas separationmembrane. In some embodiments, the functionalized PFPE network is usedas a gas separation membrane to separate gases selected from the groupconsisting of CO₂, methane, hydrogen, CO, CFCs, CFC alternatives,organics, nitrogen, methane, H₂S, amines, fluorocarbons, fluoroolefins,and O₂. In some embodiments, the functionalized PFPE network is used toseparate gases in a water purification process. In some embodiments, thegas separation membrane includes a stand-alone film. In someembodiments, the gas separation membrane includes a composite film.

In some embodiments, the gas separation membrane includes a co-monomer.In some embodiments, the co-monomer regulates the permeabilityproperties of the gas separation membrane. Further, the mechanicalstrength and durability of such membranes can be finely tuned by addingcomposite fillers, such as silica particles and others, to the membrane.Accordingly, in some embodiments, the membrane further includes acomposite filler. In some embodiments, the composite filler includessilica particles.

XV. Applications of Solvent Resistant Low Surface Energy Materials

According to alternative embodiments, the presently disclosed materialsand methods can be combined with and/or substituted for, one or more ofthe following materials and applications.

According to one embodiment, the materials and methods of the presentinvention can be substituted for the silicone component in adhesivematerials. In another embodiment, the materials and methods of thepresent invention can be combined with adhesive materials to providestronger binding and alternative adhesion formats. An example of amaterial to which the present invention can be applied includesadhesives, such as a two part flowable adhesive that rapidly cures whenheated to form a flexible and high tear elastomer. Adhesives such asthis are suitable for bonding silicone coated fabrics to each other andto various substrates. An example of such an adhesive is, DOW CORNING®Q5-8401 ADHESIVE KIT (Dow Corning Corp., Midland, Mich., United Statesof America).

According to another embodiment, the materials and methods of thepresent invention can be substituted for the silicone component in colormasterbatches. In another embodiment, the materials and methods of thepresent invention can be combined with the components of colormasterbatches to provide stronger binding and alternative bindingformats. Examples of a color masterbatch suitable for use with thepresent invention include, but are not limited to, a range of pigmentmasterbatches designed for use with liquid silicone rubbers (LSR's), forexample, SILASTIC® LPX RED IRON OXIDE 5 (Dow Corning Corp., Midland,Mich., United States of America).

According to yet another embodiment, the materials and methods of thepresent invention can be substituted for liquid silicone rubbermaterials. In another embodiment, the materials and methods of thepresent invention can be combined with liquid silicone rubber materialsto provide stronger binding and alternative binding techniques of thepresent invention to the liquid silicone rubber material. Examples ofliquid silicone rubber suitable for use or substitution with the presentinvention include, but are not limited to, liquid silicone rubbercoatings, such as a two part solventless liquid silicone rubber that isboth hard and heat stable. Similar liquid silicone rubber coatings showparticularly good adhesion to polyamide as well as glass and have aflexible low friction and non-blocking surface, such products arerepresented by, for example, DOW CORNING® 3625 A&B KIT. Other suchliquid silicone rubber includes, for example, DOW CORNING® 3629 PART A;DOW CORNING® 3631 PART A&B (a two part, solvent free, heat-cured liquidsilicone rubber); DOW CORNING® 3715 BASE (a two part solventlesssilicone top coat that cures to a very hard and very low frictionsurface that is anti-soiling and dirt repellent); DOW CORNING® 3730 A&BKIT (a two part solventless and colorless liquid silicone rubber withparticularly good adhesion to polyamide as well as glass fabric);SILASTIC® 590 LSR PART A&B (a two part solventless liquid siliconerubber that has good thermal stability); SILASTIC® 9252/250P KIT PARTS A& B (a two part, solvent-free, heat cured liquid silicone rubber;general purpose coating material for glass and polyamide fabrics; threegrades are commonly available including halogen free, low smoketoxicity, and food grade); SILASTIC® 9252/500P KIT PARTS A&B; SILASTIC®9252/900P KIT PARTS A&B; SILASTIC® 9280/30 KIT PARTS A & B; SILASTIC®9280/60E KIT PARTS A & B; SILASTIC® 9280/70E KIT PARTS A & B; SILASTIC®9280/75E KIT PARTS A & B; SILASTIC® LSR 9151-200P PART A; SILASTIC® LSR9451-1000P; RTV Elastomers (Dow Corning Corp., Midland, Mich., UnitedStates of America); DOW CORNING® 734 FLOWABLE SEALANT, CLEAR (a one partsolventless silicone elastomer for general sealing and bondingapplications, this silicone elastomer is a flowable liquid that is easyto use and cures on exposure to moisture in the air); DOW CORNING®Q3-3445 RED FLOWABLE ELASTOMER; (a red, flowable one part solventlesssilicone elastomer for high temperature release coatings, typically thisproduct is used to coat fabric, release foodstuffs, and is stable up to260° C.); and DOW CORNING® Q3-3559 SEMIFLOWABLE TEXTILE ELASTOMER (asemi-flowable one part solventless silicone elastomer).

According to yet another embodiment, the materials and methods of thepresent invention can be substituted for water based precured siliconeelastomers. In another embodiment, the materials and methods of thepresent invention can be combined with water based silicone elastomersto provide the improved physical and chemical properties describedherein to the materials. Examples of water based silicone elastomerssuitable for use or substitution with the present invention include, butare not limited to, water based auxiliaries to which the presentinvention typically applies include DOW CORNING® 84 ADDITIVE (a waterbased precured silicone elastomer); DOW CORNING® 85 ADDITIVE (a waterbased precured silicone elastomer); DOW CORNING® ET-4327 EMULSION(methyl/phenyl functional silicone emulsion providing fiber lubrication,abrasion resistance, water repellency and flexibility to glass fabric,typically used as a glassfiber pre-treatment for PTFE coatings); and DowCorning 7-9120 Dimethicone NF Fluid (a new grade of polydimethylsiloxanefluid introduced by Dow Corning for use in over-the-counter (OTC)topical and skin care products).

According to yet another embodiment, the materials and methods of thepresent invention can be substituted for other silicone based materials.In another embodiment, the materials and methods of the presentinvention can be combined with such other silicone based materials toimpart improved physical and chemical properties to these other siliconebased materials. Examples of other silicone based materials suitable foruse or substitution with the present invention include, but are notlimited to, for example, United Chemical Technologies RTV silicone(United Chemical Technologies, Inc., Bristol, Pa., United States ofAmerica) (flexible transparent elastomer suited forelectrical/electronic potting and encapsulation); Sodium Methylsiliconate (this product renders siliceous surfaces water repellent andincreases green strength and green storage life); Silicone Emulsion(useful as a non-toxic sprayable releasing agent and dries to clearsilicone film); PDMS/a-Methylstyrene (useful where temporary siliconecoating must be dissolved off substrate); GLASSCLAD® 6C (United ChemicalTechnologies, Inc., Bristol, Pa., United States of America) (ahydrophobic coating with glassware for fiberoptics, clinical analysis,electronics); GLASSCLAD® 18 (a hydrophobic coating for labware,porcelain ware, optical fibers, clinical analysis, and light bulbs);GLASSCLAD® HT (a protective hard thin film coating with >350° C.stability); GLASSCLAD® PSA (a high purity pressure sensitive adhesivewhich forms strong temporary bonds to glass, insulation components,metals and polymers); GLASSCLAD® SO (a protective hard coating fordeposition of silicon dioxide on silicon); GLASSCLAD® EG (a flexiblethermally stable resin, gives oxidative and mechanical barrier forresistors and circuit boards); GLASSCLAD® RC (methylsilicone with >250°C. stability, commonly used as coatings for electrical and circuit boardcomponents); GLASSCLAD® CR (silicone paint formulation curing to aflexible film, serviceable to 290° C.); GLASSCLAD® TF (a source of thickfilm (0.2-0.4 micron) coatings of silicon dioxide, converts to 36%silicon dioxide and is typically used for dielectric layers, abrasionresistant coatings, and translucent films); GLASSCLAD® FF (a moistureactivated soft elastomer for biomedical equipment and optical devices);and UV SILICONE (UV curable silicone with refractive index (R.I.)matched to silica, cures in thin sections with conventional UV sources).

According to still further embodiments of the present invention, thematerials and methods of the present invention can be substituted forand/or combined with further silicone containing materials. Someexamples of further silicone containing materials include, but are notlimited to, TUFSIL® (Specialty Silicone Products, Inc., Ballston Spa,N.Y., United States of America) (developed by Specialty Siliconesprimarily for the manufacture of components of respiratory masks,tubing, and other parts that come in contact with skin, or are used inhealth care and food processing industries); Baysilone Paint Additive TP3738 (LANXESS Corp., Pittsburgh, Pa., United States of America) (a slipadditive that is resistant to hydrolysis); Baysilone Paint Additive TP3739 (compositions that reduce surface tension and improve substratewetting, three acrylic thickeners for anionic, cationic, nonionic andamphoteric solutions, such as APK, APN and APA which are powderedpolymethacrylates, and a liquid acrylic thickener); Tego Protect 5000(Tego Chemie Service GmbH, Essen, Germany) (a modifiedpolydimethylsiloxane resin typically for matte finishes, clear finishesand pigmented paint systems); Tego Protect 5001 (a silicone polyacrylateresin that contains a water repellent, typically used with clear varnishsystems); Tego Protect 5002 (a silicone polyacrylate resin that can berepainted after mild surface preparation); Microsponge 5700 Dimethicone(a system based on the Microsponge dimethicone entrapment technologywhich is useful in the production of emulsion, powder, and stickproducts for facial treatments, foundations, lipsticks, moisturizers,and sun care products, dimethicone typically is packed into the emptyspaces in a complex crosslinked matrix of polymethacrylate copolymer);350 cST polydimethylsiloxane makes up 78% of the entrapped dimethiconecomponent and 1000 cST polydimethylsiloxane constitutes the other 22%,the system typically facilitates the delivery of dimethicone'sprotective action to the skin); MB50 high molecular weightpolydimethylsiloxane additive series (enables better processing withreduced surface friction and faster operating speeds, commonly availablein formulations for PE, PS, PP, thermoplastic polyester elastomer, nylon6 and 66, acetal and ABS, the silicone component is odorless andcolorless and can be used for applications involving food contact, theproduct can be used as a substitute for silicone fluid and PTFE);Slytherm XLT (a new polydimethylsiloxane low temperature heat transferfluid from Dow Corning, unlike traditional organic transfer fluids, itis non-toxic, odorless and does not react with other materials in thesystem, at high temperatures it has the additional advantage of beingnon-fouling and non-sludge forming); and 561® silicone transformer fluid(this material has a flash point of 300° C. and a fire point of 343° C.,the single-component fluid is 100% PDMS, contains no additives, isnaturally degradable in soils and sediments, and does not cause oxygendepletion in water).

XVI. Applications to Devices

The dual curable component materials of the present invention can beused in various medical applications including, but not limited to,medical device or medical implants. According to an embodiment, materialincluding one or more blended photo curing and/or one or more thermalcuring components can be used to fabricate medical devices, devicecomponents, portions of devices, surgical devices, tools, implantablecomponents, and the like. The blended material refers to the mixing ofphotocurable and/or thermally curable constituents within the polymerthat is to form the device. The use of such a system allows for theformation of discrete objects by activating the first curing system andthen, in some embodiments, adhering such discrete objects to otherobjects, surfaces, or materials by activating the second curing system.In some embodiments, the dual cure materials can be used to make amedical device or implant outside the body through a first cure of thematerial, then the second cure can be utilized to adhered the device totissue after implantation into the body. In other embodiments, the dualcure materials can be used to make medical devices or implants in stagesand then the components can be cured together to form an implant. Themedical devices or implants made with the dual cure materials of thepresent invention can be, but are not limited to, orthopedic devices,cardiovascular devices, intraluminal devices, dermatological devices,oral devices, optical devices, auditory devices, tissue devices, organdevices, neurological devices, vascular devices, reproductive devices,combinations thereof, and the like. Other devices and components thatcan be coated with the materials and methods disclosed herein include:tubes, piping; vials, glassware, storage containers, transfercontainers, valves, pipettes; nozzles; labware, goggles, glasses,gloves, sinks, tanks (such as fuel tanks), cylinders, ceramic devices,metal devices, polymer devices, glass, combinations thereof, and thelike. The materials can be used as coating or linings for such devicesto prevent fouling, prevent contamination, prevent reactions between thecontainer and a substance in the container, ease of cleaning,combinations thereof, and the like. In some embodiments, the materialsof the present invention adhere to these devices and materials throughhydroxy groups on the surface of the devices and form stronginteractions which maintain the coating and linings in position.

XVI.A. Forming Devices or Implants and Attaching Same to Another Deviceor Implant

According to some embodiments, an object, such as a component of amedical device or an implant, or the entire device can be fabricated byforming a liquid precursor in a mold, activating a curing mechanism,such as photo curing or thermal curing to solidify or partially solidifythe precursor, and removing the solid object from the mold. The objectcan then be placed in contact with another component of a medical deviceor implant, a surface, a coating, or the like and a second curingmechanism is activated, such as thermal curing or photo curing, toadhere the two objects together. For example, the object can be, but isnot limited to, an artificial joint component, an artificial bonecomponent, an artificial tooth or tooth component, an artificialarticular surface, an artificial lens, and the like.

An example of this procedure is shown in FIG. 13, steps A-C, forexample. According to FIG. 13, liquid fluoropolymer (e.g., PFPE)material can be introduced into a mold and upon activating a firstcuring mechanism (e.g., thermal curing) the PFPE material is solidifiedto form a tube 1300. Tube 1300 can then be inserted into a second tube1310, formed from the same or a different material, such as for examplea different polymer or a natural material or structure such as tissue ora blood vessel. Next, a second curing mechanism (e.g., photo curing) isactivated to adhere the PFPE tube 1300 to the second tube 1310.

According to some embodiments directed to orthopedic applications, thedual curable materials of the present invention facilitate rebuildingand/or building new devices and structures for placement within a livingbody. Further embodiments include rebuilding and repairing existingbiologic or artificial devices, tissues, and structures in situ. Forexample, dual curable materials may be utilized in building new jointsand in repairing existing joints in vitro or in situ.

According to some embodiments, a damaged biologic component can be adamaged tissue such as skeletal tissue (e.g., spinal components such asdiscs and vertebral bodies, and other skeletal bones). In someembodiments, the dual cure materials of the present invention can beused in situ to augment the damaged biologic components. According tosuch embodiments, the method includes surgically inserting a moldstructure into the damaged site or preparing the surgical site to act asa receiving mold for liquid dual cure material. The mold structure isconfigured to receive liquid dual curable material and is geometricallyconfigured similar to the damaged biologic component to be replaced orconfigured to yield a desired result. Next, liquid dual cure material isintroduced into the mold and first cured. The first cure can be, forexample, treatment with light or heat. In some embodiments, the firstcure can be an incomplete cure such that the replacement structure isleft compliant. The compliant nature of the replacement structure canfacilitate removal of the mold structure from the site of damage orpositioning of the replacement component in the desired surgical/implantsite. After removal of the mold structure, the first cured replacementstructure can be treated with a second cure to further cure thereplacement structure to satisfy desired mechanical properties for theparticular application.

In other embodiments, the replacement component can be built up, eitherin vitro or in situ. According to these embodiments, an opening to asurgical site may be made smaller than an implant required by the sitebecause the implant can be built up a portion at a time (e.g., replacinga hip joint through an arthroscopic type procedure). In suchembodiments, dual cure liquid material, as described herein, can beintroduced into a mold or a surgical site and treated with a first cure.The first cure activates the liquid material to form a first portion ofthe replacement component such that the component can retain a desiredshape. The first portion can be configured to harden upon the firstcuring or remain compliant. Next, in some embodiments, a second quantityof liquid material can be introduced to form a second portion of thereplacement component. The material of the second portion is treatedwith a first cure treatment. The first cure treatment used to treat thesecond portion of the component can be the same technique used on thefirst portion such that each component retains a viable second curecomponent. Therefore, because each portion of the device retains aviable second cure component, after the first cured portions of thedevice are compiled to form the completed device, the portions can betreated with a second curing. Upon second curing, the second curecomponent of the layered portions of the device will be activated andthe layered portion will bind together forming one integral device.Multiple portions of the replacement component can be formed, asdescribed herein, as needed to make a replacement device. According tosome embodiments, each portion can have different functional and/ormechanical properties to impart a desired mechanical and/or chemicalresult on the completed replacement component.

According to other embodiments, a portion such as an articular surfaceof a joint can be formed with the dual cure material of the presentinvention and attached to a natural joint in situ. According to suchembodiments, an artificial articular surface can be fabricated from afirst cure (e.g., thermal cure) of the dual cure material of the presentinvention. The artificial articular surface can then be implanted onto apreexisting joint surface and treated with a second cure (e.g., photocure) such that the artificial articular surface binds to thepreexisting joint surface.

XVI.B. Forming Devices or Implants and Attaching Same to Tissue

According to other embodiments, dual cure materials of the presentinvention can be used to replace or augment natural biologic tissue orstructures and can be adhered directly to the tissue.

According to some embodiments, dual cure materials described herein maybe incorporated into various types of repairs or patches, as shown inFIG. 14. In one embodiment, such a patch can be utilized in lungsurgical procedures. Patches include, for example, but are not limitedto sheets of dual cure material configured to be attached and secureddirectly to living tissue through activation of the second curingmechanism of the dual cure materials.

According to some embodiments, a disrupted or damaged material, device,or surface can be repaired (e.g., patched) with material of the presentinvention. As shown in FIG. 14, steps A-C, a patch can be made bymolding dual cure material into a desired shape and activating a firstcure (e.g., thermal cure) to form patch 1400. Next, patch 1400 is placedover a device or tissue 1410 that is affected with a disruption ordamage (e.g., a crack, hole, surgically altered tissue) 1412. Afterplacement of patch 1400 over disruption 1412, a second curing mechanismis activated (e.g., photo curing) to adhere the patch to the surface ofdevice 1410. The strength of the patch is dependent upon multiplevariables, such as, for example, the size of binding area between patch1400 and tissue 1410, the extent of curing administered to thepatch/tissue combination, the chemicals, quantities, concentrations, andthe like used in the second curing process, the composition of patch1400, the composition of tissue or device 1410, combinations thereof,and the like. According to alternative embodiments, patch 1400 canundergo a second cure (e.g., thermal or photo curing) to attach patch1400 to a compound, material, or substance that is known to bind totissue. For example, patch 1400 can be treated with or adhered to afribrin sealant component or glue, which is well known and usedextensively in various clinical settings for adhering tissues together.In other embodiment, the patch can be second cure attached to abiocompatible material and the biocompatible material is then stitchedto the tissue, thereby implanting the patch.

In yet other embodiments, the materials of the present invention can beused to fabricate a mold and replicate another object. In someembodiments the object to be molded and replicated can be a medicaldevice or a tissue, such as a joint component, organ, organ scaffold,joint, skeletal component, dental component, ocular component, vascularcomponent, and the like.

According to such embodiments, as shown in FIG. 15, steps A-E, a mold isfabricated by taking an object such as a bone 1500 and encapsulating theobject in a curable matrix 1502 such as liquid PDMS precursors. Next,the curable matrix 1502 is cured. The bone 1500 is then removed, leavinga mold 1504 in a shape that corresponds to the molded object 1500. Insome embodiments, the cured mold can be reversibly swelled to assist inremoval of the object. Next, mold 1502 can be filled with dual curematerials of the present invention, such as for example, dual cureliquid PFPE precursors 1510. The dual cure material 1510 is thensubjected to a first cure (e.g., thermal curing) to form a replicateobject 1512 in the shape of bone 1500. Next, the replicate object 1512can be implanted into the body as a replacement component. Duringimplantation replicate object 1512 can be adhered to natural tissues,such as articular cartilage, portions of remaining natural bone,ligaments, tendons, other artificial joint components, and the like, bypositioning the tissues with respect to replicate object 1512 andsubjecting the combination to a second cure (e.g., photo curing).

According to other embodiments, dual cure materials of the presentinvention can be used in various cardiovascular applications and inother intraluminal applications. In some of these embodiments, thematerials can be used to fabricate and/or augment body lumens, and toform artificial lumens (e.g., artificial blood vessels). The dual curematerials of the present invention can be molded, as shown in FIG. 15,to form replacement blood vessels for replacing damaged and/or occludedvessels within a body. Not only can the materials disclosed herein serveas conduits for blood flow, but they also can allow for diffusion ofoxygen and nutrients through the vessel wall into surrounding tissuesthus functioning much like a normal healthy blood vessel.

According to embodiments of the present invention, a method ofreplacing, in situ, a portion of a blood vessel includes injecting anoxygen permeable, bacterial impermeable dual cure liquid PFPE materialinto a lumen of a portion of a blood vessel such that the dual cureliquid PFPE coats the luminal surface of the blood vessel. The dual cureliquid PFPE is then subjected to a first cure technique to form anartificial blood vessel within the natural blood vessel. The biologicblood vessel can then be removed from the first cured PFPE material andthe material can be subjected to a second cure or can be treated withanother layer of dual curable liquid PFPE and the combination can besubjected to a second cure. Also, the second cure can be applied whenthe artificial blood vessel is positioned within the subject andactivated to bind the material to the natural blood vessel. A workingreplacement for the blood vessel portion is thereby produced.

Embodiments of the present invention are particularly advantageousregarding repair and/or replacement of blood vessels. Given their highoxygen carrying ability and permeability, artificial vessels formed fromPFPE materials have highly functional properties with syntheticvasavasorum characteristics. PFPE materials allow diffusion of oxygenthrough the walls and into surrounding dependent tissues, allowdiffusion of sustaining nutrients, and diffusion of metabolites. PFPEmaterials substantially mimic vessels mechanically as they are flexibleand compliant. Moreover, embodiments of the present invention areparticularly suitable for use in heart by-pass surgery and as artificialarterio-venous shunts. PFPE materials can also be used to repair naturalor synthetic arterio-venous shunts by coating the inside surface of thedamaged or worn vessel and curing as described herein. According toother embodiments, intraluminal prostheses can be employed in sites of abody such as, but not limited to, biliary tree, esophagus, bowel,tracheo-bronchial tree, urinary tract, and the like.

In another embodiment, the dual cure materials of the present inventioncan be used to fabricate stents for repairing vascular tissue. In someembodiments the dual cure liquid material can be locally implanted andcured during a balloon angioplasty procedure, or the like, and subjectedto a curing or a second curing after being locally positioned. In suchembodiments, the dual cure liquid material can be first cured to form amanipulable sheet or tube of material. The manipulability of the sheetfacilitates implantation of the stent precursor material. The stentprecursor material can then be positioned, for example, by anangioplasty procedure. Upon positioning the stent precursor theimplantation device can subject the stent precursor material to a secondcuring, thereby, creating a mechanically viable stent.

The dual cure PFPE materials, according to embodiments of the presentinvention, may be used with all of the cardiovascular and intraluminaldevices described herein. PFPE materials may be utilized in thematerial(s) of these devices and/or may be provided as a coating onthese devices.

According to other embodiments and as shown in FIGS. 16A-16C, a biologicstructure having a lumen (e.g., for example, a blood vessel) can bereplaced with a medical device molded from the dual cure materialsdisclosed herein. FIG. 16 shows a top view or end view of a biologicstructure 1602 having a lumen 1604. First, in molding the replacementvessel, lumen 1604 is filled with a temporary filler 1603. Temporaryfiller 1603 can be PDMS, foam, or another suitable material that can beinserted into vessel 1602. Filler 1603 can be administered into lumen1604 such that a desired pressure is applied to the walls of vessel1602. The pressure applied to the walls of vessel 1602 can be a pressureto mimic a biologic condition, a pressure below a normal biologiccondition, a pressure above a normal biologic condition, a desiredpressure, or the like. Vessel 1602 is then encapsulated into curableouter matrix 1600, such as for example liquid PDMS. Next, outer matrix1600 is cured such that vessel 1602 is sandwiched between outer matrix1600 and filler 1603.

Referring now to FIG. 16B, vessel 1602 is removed from between outermatrix 1600 and filler 1603, creating a receiving space 1606. Next,replacement material (e.g., liquid PFPE or the like) having dual curecapabilities, as described herein, is delivered into receiving space1606. Replacement material can be injected, poured, sprayed, or the likeinto receiving space 1606. Next, replacement material is subjected to afirst cure (e.g., photo or thermal curing) such that it solidifies, atleast partially, and forms replacement device 1620. Following the firstcure, outer matrix 1600 and filler 1603 are removed, thereby, leavingreplacement device 1620 (FIG. 16C). Replacement device 1620 has an outersurface 1610, an inner surface 1612, and includes the characteristics ofthe natural biologic structure from which it was molded. Furthermore,replacement device 1620 includes a lumen 1608 that mimics the lumen ofthe biologic structure that replacement device 1620 was molded from.

Next, replacement device 1620 is positioned into the subject in theposition where the natural component was removed or any other suitableimplant location. Replacement device 1620 is aligned with biologicstructures that it is configured to adhere to and function with andreplacement device 1620 is treated with a second cure (e.g., thermal orphoto curing). The second cure activates components of the replacementdevice (described herein) which in turn bind with the surroundingbiologic tissue, thereby implanting and affixing replacement device 1620with the subject. In other embodiments, replacement device 1620 can bebound to a bio-active polymer that is known to adhere to tissue, in thesecond curing step, such that the replacement device 1620 can bind tothe biologic tissue through the bioactive polymer.

According to another embodiment, the dual cure material can be used toform a rigid structure that augments structural support to a skeletalportion of the subject. For example, damage to be augmented can be acrack or other defect in a bone. In some embodiments, the dual cureliquid material can be first molded or formed in vitro and first curedto form a structure of desired configuration. Next, the first curedstructure can be implanted and positioned with respect to the damagedbiologic structure to be augmented. Once in position, the first curedmaterial can be treated with a second cure to further solidify and/orbond to the biologic structure. The dual cure mechanism of the presentinvention facilitates implantation of the structure because upon firstcuring the structure can retain a specific shape but be very compliant.The compliant nature of the structure after the first cure can reducetrauma inflicted on a patient while implanting the structure. Upon thesecond curing, the structure binds with the adjacent tissue or biologiccomponent, seals the crack, and provides structural support to thedamaged biological component. The composition and degree of curing ofthe implanted material can be altered to render a structure thatresembles a desired functionality, such as strength, flexibility,rigidity, elasticity, combinations thereof and the like. Accordingly,the dual cured, flexible material may replace portions of ligaments,tendons, cartilage, muscles, and the like as well as tissue (e.g.,flexible tissues) within the body of a subject.

In still further embodiments, the dual cure materials of the presentinvention can be utilized to form other medical devices, medicalimplants, biological replacement devices, medical procedure tools,surface treatments, combinations thereof, and the like. Other usefulapplications to which the dual cure material can be applied aredisclosed in published U.S. patent application no. 2005/0142315,including the publications cited therein, all of which are incorporatedherein by reference in their entirety.

In still further embodiments, the dual cure materials disclosed anddescribed herein can be used to form a patterned surface characteristicon the surface of medical devices. The patterned surface characteristiccan provide useful properties to medical devices and as medical devicecoatings. The surface patterning of medical devices and medical implantscan provide superhydrophobic coatings that can be extremely non-wettingto fluids. The patterned surfaces can also be highly resistant tobiological fouling. Dual cure materials can be patterned by pouring aliquid precursor of the dual cure material onto a patterned template(e.g., silicon wafer) or by photolithography, and treating the precursorto a first curing, whereby the material solidifies or partiallysolidifies and takes the shape of the pattern on the patterned wafer. Insome embodiments, the pattern can have structures that are between about1 nm and about 500 nm. In other embodiments, the pattern can havestructures that are between about 1 μm and 10 μm. In one embodiment thepattern is a repeated diamond shape pattern.

Next, the first cured material is released from the wafer to yield apatterned layer. Such a layer can then be used directly as a medicaldevice or can be adhered, through a second curing, to other objects bythe orthogonal curing methods previously described, thereby coating thesurface of medical devices and implants and resulting in decreasedwettability and decreased likelihood of bio-fouling of the medicaldevice or implant.

In other embodiments, the dual cure materials are useful indermatological applications including, for example, bandages, dressings,wound healing applications, burn care, reconstructive surgery, surgicalglue, sutures, and the like. Because PFPE materials are oxygen permeableand bacterial impermeable, tissue underlying a PFPE bandage can receiveoxygen while being protected against the ingress of dirt, microbialorganisms, pathogens, and other forms of contamination and toxicity. Inaddition, the oxygen permeability and carrying capacity of PFPEmaterials can also help with preventing necrosis of healthy tissue underbandages and dressings, or under an area being treated.

According to an embodiment of the present invention, a method ofapplying “instant skin” to the body of a subject includes applying anoxygen permeable, bacterial impermeable liquid dual cure PFPE materialonto a portion of the body of a subject. The dual cure PFPE material canbe treated with a first cure to form layers of an approximatepredetermined size and/or shape. After the first cure, the layered PFPEdual cure material is placed on the damaged zone of the patient. Thedual cure PFPE is then subjected to a second cure such that the dualcure PFPE adheres to the patient and provides a oxygen permeable,microbial impermeable, waterproof, flexible, elastic, biocompatibleartificial skin layer.

According to further embodiments, ocular implants and contact lenses canbe formed from the dual cure materials of the present invention. Thesedevices are advantageous over conventional ocular implants and contactlenses because the PFPE material is permeable to oxygen and resistant tobio-fouling. In addition, because of the lower surface energy, there ismore comfort to the wearer as a result of the low friction generated thePFPE. In addition, the refractive index of PFPE materials can beadjusted for optimum performance for ocular implants and contact lenses.Further embodiments include cochlear implants utilizing the dual curePFPE material. Using dual cure PFPE materials, tissue in-growth can beminimized, thus making removal of the device safer and less traumatic.

XVII. Materials Having Nanoscopic Voids and Methods for Forming the Same

According to other embodiments of the present invention, materials ofthe present disclosure are formed with nano-scale voids. The nano-scalevoids can provide a porous material and/or increase the permeability ofthe material. According to such embodiments, a fluorinated ornon-fluorinated fluid is introduced to the precursors, described herein,in low concentrations. The materials are then cured as described herein,including but not limited to UV curing, thermal curing, evaporation,combinations thereof, and the like. Next, the fluid is evaporated orremoved from the cured material. Following removal of the fluid from thecured material, nano-scopic voids are left behind. These nano-scalevoids can give porosity to the material, increase permeability of thematerial, can be interconnected or independent, combinations thereof,and the like. According to one embodiment, the concentration of thefluid is less than about 15%. According to another embodiment, theconcentration of the fluid is less than about 10%. In yet anotherembodiment, the concentration of the fluid is less than about 5%.According to such embodiments, the fluid acts as a porogen, leavingnano-scopic voids in the cured elastomer, thereby increase the gaspermeability of the material, generating nano-scale porosity in thematerial, increasing liquid permeability, combinations thereof, and thelike. Other methods for fabricating nanoscopic voids in the materials ofthe present invention are disclosed in U.S. Pat. No. 6,160,030 to Chaouket al., which is incorporated herein by reference in its entiretyincluding all references cited therein.

XVIII. Other Applications

According to other embodiments of the present invention, traditionalapplications for silicone can be improved with the materials and methodsof the present invention and according to further embodiments theapplications can be replaced with the materials and methods disclosedherein. Silicone applications to which the materials and methods of thepresent invention are applicable include mold release agents, releaselayers, respiratory masks, anti-graffiti paint systems; aqueouscoatings, sealants, mechanically assembled monolayers, micro plates andcovers, tubing, water repellant, and organic solvent repellant.

Microextraction is a further application to which the materials andmethods of the present invention can be applied. For example, thematerials and methods of the present invention can be applied tosubstitute for or enhance the current techniques and chemicals used inmicroextraction. An example of microextraction is detailed in an articlein Analytical Chemistry [69 (6), 1197-1210, 1997] in which the authorsplaced 80 microliter chips of OV-1 extraction medium[poly(dimethylsiloxane)] in 50 ml flasks with 49 ml of aqueous sample,shook the flasks for 45 to 100 minutes, removed the chips, and placedthem in the cell of a Shimadzu UV-260 spectrophotometer (Shimadzu Corp.,Kyoto, Japan) to obtain a UV spectrum. Further described is apreconcentration by SPME that enables UV absorption spectroscopy toidentify benzene at detection limits of 97 ppb, naphthalene at 0.40 ppb,1-methylnaphthalene at 0.41 ppb, and 8 other aromatics at 5.5-12 ppb. Intests of samples spiked with unleaded gasoline, JP4 jet fuel, and no. 1diesel fuel, preconcentration permits direct quantitation of dilutelevels of aromatic species in aqueous samples without interference fromhumic substances in solution.

According to other embodiments, an application of the present inventioncan include substituting the materials and methods of the presentinvention for traditional chromatographic separation materials.According to yet another embodiment of the present invention thematerials and methods of the present invention can be combined withtypical chromatographic separation materials. Chromatographicseparations useful with the present invention are described in thefollowing studies, incorporated herein by reference, and which describethat natural enantiomeric distribution of terpene alcohols on variousnatural matrices determined that, although distinctive for each matrix,the distribution is widely differentiated. While there is data availableon the free bound linalool content in muscat wines, no data is availableon the enantiomeric distribution of the same terpene alcohols in thesewines. Researchers at DIFCA (Sezione di Chimica AnaliticaArgoali-mentare ed. Ambientale, Universita degli Studi di Milano, ViaCeloria 2, 20133 Milan, Italy; Tel: 39 2 26607227, Fax: 39 2 2663057)have characterized muscat wines using gas chromatography (GC) chiralanalysis. To determine the aromatic fraction of muscat wines, theenantiomeric excess of linalool and α-terpineol must be measured. F.Tateo and M. Bononi used two different fibers for the solid phasemicroextraction (SPME), one apolar (100 micron non-bondedpolydimethylsiloxane) and one polar (partially crosslinked 65 microncarbowax/divinylbenzene). There was greater adsorption of the linaloolusing the polar fiber. The enantiomeric distribution of the linalool andof the α-terpineol were within fairly narrow limits and were consideredcharacteristic indices. In order to assess the selectivity of SPMEadsorption of the polar fiber with respect to a number of molecules,comparison was made using data obtained by direct injection. Greatersensitivity for the molecules was obtained using this technique.

In other applications, the materials and methods of the presentinvention can be substituted for PDMS materials used in outdoorcapacities, such as for example, PDMS shed materials used to cover highvoltage outdoor insulators. According to further embodiments, thepresently disclosed materials and methods can be combined with PDMSmaterials used in outdoor capacities, such as for example, the highvoltage outdoor insulator sheds described above. It is important thatthe surface of the shed of the insulator remains hydrophobic throughoutits services life. It is known, however, that electrical discharges leadto an oxidation of the traditional surface and a temporary loss ofhydrophobicity. According to a study of traditional materials,crosslinked polydimethylsiloxane (PDMS) containing Irganox 1076, Tinuvin770 or Irganox 565 (Ciba Specialty Chemicals Corp., Tarrytown, N.Y.,United States of America), prepared by swelling PDMS in a solution ofone of these stabilizers in n-hexane, was exposed to a corona dischargeand the corona exposure time (t-crit) to form a brittle, silica-likelayer was determined by optical microscopy. The critical corona exposuretime showed a linear increase with increasing stabilizer concentration;Tinuvin 770 showed the highest efficiency and Irganox 1076 the lowest.The increase in t-crit on corona exposure of the stabilized samples withreference to the value for unstabilized PDMS was similar to thatreported earlier for air plasma exposed samples. The efficiency of thestabilizers towards corona-induced surface oxidation of PDMS also wasconfirmed by X-ray photoelectron spectroscopy. As will be appreciated byone of ordinary skill in the art, however, traditional materialsutilizing PDMS can be significantly improved by the addition oraugmentation with the materials and methods of the present invention.

Microvalves actuated by paraffin, such as, for example, microvalvescontaining silicone-rubber seals actuated by heating and cooling ofparaffin have been proposed for development as integral components ofmicrofluidic systems. According to an embodiment of the presentinvention, the materials and methods of the present invention can besubstituted for, or combined with the silicone-rubber seals of suchdevices as the disclosed microvalve materials, thereby, increasing thephysical and chemical properties of such microvalves.

Scratch-free surfaces are yet a further application to which thematerials and methods of the present invention can be applied. Thematerials and methods of the present invention can be substituted for orused to augment the traditional scratch-free surface materials toimprove their physical and chemical properties. As an example, researchfrom Dow Corning of Freeland, Mich., has shown that adding masterbatchesto thermoplastic olefins (TPOs) improves scratch resistance of TPOcomponents. The company's MB50-series of masterbatches are incarrier-resin formats containing 50% ultra-high molecular weightpolydimethylsiloxane, a scratch-resisting and lubricating additive. Theadditive lowers the coefficient of friction at the surface of the moldedpart. Surface-modifying masterbatches are now utilized and developed forvarious applications, such as in the automotive sector where it is beingused in consoles, airbags, door skins and exterior components.Substituting the materials and methods of the present invention, orcombining the materials and methods of the present invention to suchscratch-free surface materials can improve the scratch-resistance of thematerials.

The materials and methods of the present invention also can be appliedto materials and methods used in the fabrication of sensors. An exampleof applying the materials and methods of the present invention to thematerials that are used to make sensors, such as for example, polymericmembrane paste compositions will be appreciated by one of ordinary skillin the art from the following. Advantageous polymeric membrane pastecompositions include a polyurethane/hydroxylated poly(vinyl chloride)compound and a silicone-based compound in appropriate solvent systems toprovide screen-printable pastes of the appropriate viscosity andthixotropy. For an ion sensor to be commercially acceptable, it musthave qualities beyond just electrochemical performance. For a sensor tobe cost effective, it must be reproducible using mass productionsystems. There must be common electrochemical response characteristicswithin the members of a batch fabricated group. If the sensors are notall substantially identical, they will each be characterized bydifferent lifetimes and response characteristics, creating difficultiesin the field, not the least of which is the added cost associated withrecalibration of equipment whenever the sensor is changed. Polymericmembranes are in common use as transducers in solid-state chemicalsensors, particularly because such membranes have high selectivity tothe ion of interest and can be made selective to a wide range of ionsusing one or many readily available ionophores. One known technique forforming the membranes is solvent casting; a technique which originatedwith ion-selective electrode technology. In addition to being a rathertedious operation, particularly in view of the small size of thesensors, this production method yields very high losses. The thicknessand shape of the membrane cannot be controlled, resulting in anunacceptable lack of sensor reproducibility. An objective of theresearch at the University of Michigan was to provide a simple andeconomical system for batch fabrication of solid-state ion-selectivesensors. Their method consists of installing a mask on a semiconductorsubstrate, the mask having at least one aperture having a predeterminedconfiguration which corresponds with a desired membrane configuration. Apolymeric membrane paste is applied to the mask, and a squeegee is drawnacross the mask to force the paste into the aperture and incommunication with the semiconductor substrate. In one form, the mask isof a metallic material, which can be a stainless steel mesh coated witha photoreactive emulsion. In another form, the mask is a metal foilstencil. The membrane which ultimately is produced has a thickness whichcorresponds to that of the mask, between 25 and 250 microns. Themembrane paste can be formed of a polyurethane with an effective portionof an hydroxylated poly(vinyl chloride) copolymer; a polyimide-basedcompound; a silicone-based compound, such as silanol-terminatedpolydimethylsiloxane with the resistance-reducing additive,ON-derivatized silicone rubber; or any other suitable polymericmaterial. Thus, it will be appreciated that the materials and methods ofthe present invention can be applied to the materials and processes forforming sensors, such as the polymeric membrane compositions,polyimide-based compounds, polydimethylsiloxane, and silicone rubber,for example.

In another further embodiment, the materials and methods of the presentinvention can be substituted for or can be used to augment the materialsand methods used in sol-gel capillary microextraction. Typically,sol-gel technology involves the encapsulation of active ingredients inmicro- and nano-sized matrices, often silica based matrices, as well asnanospheres. Sol-gel capillary microextraction (sol-gel CME), forexample, is a viable solventless extraction technique for thepreconcentration of trace analytes. Sol-gel-coated capillaries are oftenemployed for the extraction and preconcentration of a wide variety ofpolar and nonpolar analytes. Two different types of sol-gel coatings areused for extraction: sol-gel poly(dimethylsiloxane) (PDMS) and sol-gelpoly(ethylene glycol) (PEG). A gravity-fed sample dispensing unit can beused to perform the extraction. The analysis of the extracted analytescan be performed by gas chromatography (GC). The extracted analytes aretransferred to the GC column via thermal desorption. For this, thecapillary with the extracted analytes can be connected to the inlet endof the GC column using a two-way press-fit fused-silica connector housedinside the GC injection port. Desorption of the analytes from theextraction capillary can be performed by rapid temperature programming(at 100 degrees C./min) of the GC injection port. The desorbed analytesare transported down the system by the helium flow and further focusedat the inlet end of the GC column maintained at 30 degrees C. Sol-gelPDMS capillaries are commonly used for the extraction of nonpolar andmoderately polar compounds (such as, but not limited to, polycyclicaromatic hydrocarbons, aldehydes, ketones), while sol-gel PEGcapillaries are used for the extraction of polar compounds (such as, butnot limited to, alcohols, phenols, amines). For both polar and nonpolaranalytes, the run-to-run and capillary-to-capillary relative standarddeviation (RSD) values for GC peak areas often remain under 6% and 4%,respectively. Parts per quadrillion level detection limits are achievedby coupling sol-gel CME with gas chromatography/flame ionizationdetection (GC-FID). The use of thicker sol-gel coatings and longercapillary segments of larger diameter (or capillaries with sol-gelmonolithic beds) often lead to further enhancement of the extractionsensitivity. As will be appreciated by one of ordinary skill in the art,replacing or combining the matrices and nanospheres commonly used insol-gel applications with the materials and methods of the presentinvention can improve the efficiency and effectiveness of sol-gelprocesses.

In alternative embodiments the materials and methods of the presentinvention can be applied to processes, such as process aid for plasticsand membrane separating processes. An example of membrane separatingprocesses applicable with the present invention is described in Membrane& Separation Technology News, v. 15: no. 6, Feb. 1, 1997(ISSN-0737-8483)).

Other silicone related arts that the methods and materials of thepresent invention are capable of augmenting or replacing include, butare not limited to, the disclosures in U.S. Pat. Nos. 6,887,911;6,846,479; 6,808,814; 6,806,311; 6,804,062; 6,803,103; 6,797,740; thedisclosures in U.S. Patent Application Nos. 2005/0147768; 2005/0112385;2005/0111776; 2005/0091836; 2005/0052754; and the disclosure inEP1533339A1, each of which are incorporated by reference herein in theirentirety.

XIX. Sacrificial Layer Device Fabrication

According to some embodiments of the present invention, the materialsdescribed herein are used to form devices, such as microfluidic devices,fabricated according to sacrificial layer methods. Specificphotolithographic patterning of curable materials disclosed herein,through controlled light exposure, such as for example, rastering afocused source, flood exposure through a mask, a highly organizedprecision light source, combinations thereof, or the like providesfabrication of polymeric micro-devices. Furthermore, according toembodiments of the present invention, living radicalphoto-polymerization of materials disclosed herein provides parallelfabrication of multilayer devices. Living radical photo-polymerization,uses specific initiator chemistries that continuously reactivate duringexposure to UV light for synthesizing linear polymers. According to someembodiments, the polymer chain ends are not permanently terminated ordeactivated and in conjunction with cross-linking monomers, livingradical photo-polymerization enables initiation and covalent adhesion ofnew polymer chains or films to surfaces of previously photo-polymerizedstructures.

According to some embodiments, methods for photo patternedpolymerization of polymers, such as polymers discloses herein, can beused to generate micro-devices, such as microfluidic devices, withhighly complex three-dimensional designs, extremely versatile surface,and a plurality of physical and chemical properties. In alternativemethods, the materials disclosed herein can be combined with materials,such as filters, different polymerizable monomer formulations, such asfor example, photo-curable agents, thermal curable agents, combinationsthereof, or the like to form cured components that can be combined aslayers of an overall structure or device. Following initial curing ofthe materials, adjacent layers or devices can be covalently adheredtogether, through a second curing process or procedure, such asdescribed herein, to form complex multilayered devices. According tosome embodiments, material properties can be initially selected byaltering the composition of the base material and/or monomer formulationin the base material, according to materials and methods describedherein, such that the resulting layers of a device or the device itselfwill function for a predetermined use.

According to FIG. 18, a reaction chamber 1800 is provided. Reactionchamber 1800 is typically shaped similar to a cup, with sides 1802 suchthat it can house a liquid 1804 without the liquid spilling from chamber1800. Chamber 1800 can have a flat, curved, or the like bottom chamberor can have a shape or recesses 1806 contained therein. Chamber 1800preferable has sides 1802 with a height equal to or greater than aheight of a desired device. In some embodiments, sides 1802 are betweenabout 1 nm and about 100 mm. According to other embodiments, sides 1802are between about 1 micron and about 500 micron. According to yet otherembodiments, sides 1802 are between about 50 nm and about 50 mm. Liquidmaterial 1804, such as materials disclosed herein, for example, PFPE ora derivative of PFPE as disclosed herein, is presented into chamber 1800and filled to a desired depth. The desired dept can be determined by adesired thickness of the resulting device or layer to be fabricated in afirst fabrication step.

According to some embodiments, a mask 1810, such as for example, anoptical mask alignment system having a desired mask design or pattern1812 is placed above or in contact with liquid polymer material 1810. Insome embodiments, masks 1810 are made from commercially availableemulsion films with a high resolution plotter. According to suchembodiments, masks 1810 are made from rapid prototyping and deviceoptimization. Using a method of mask fabricating such as these typicallygenerates a resolution of about 10 micron. According to otherembodiments, higher resolution, i.e., less than about 1 micron and/ormore durable masks can be fabricated by deposition of chrome on etchedquartz glass or alternative printing methods. Masks 1810 can be alignedusing registration marks or an optical alignment system. In someembodiments, mask 1810 and liquid material 1804 exposed by the pattern1812 of mask 1810, is treated with a light or UV cure 1830, such asdescribed herein. In some embodiments cure 1830 activates photo-curablemonomer components in liquid material 1804 such that liquid material1804 is cured in a pattern according to pattern 1812 of mask 1810.According to other embodiments, after liquid material 1804 is presentedinto chamber 1800, liquid material 1804 is treated with a highlyorganized light source such that primarily predetermined areas of liquidmaterial 1804 is cured to a solid form or a substantially solid form. Ifmask 1810 is utilized, it is then removed and any uncured liquidmaterial 1804 is removed by treating the uncured liquid material 1804with a solvent.

According to some embodiments, it can be desirable to form and adhere asecond layer or a second device to the initially cured liquid material1804. According to such embodiments, voids 1818 left behind afterremoving uncured liquid material 1804 are filled with a sacrificiallayer 1820. In some embodiments, sacrificial layer 1820 can be a wax,such as paraffin wax, or the like. Sacrificial layer 1820 is preferablya liquid or molten solid such that it fills voids 1818 to preserve openspaces in the material, such as for example, channels, valve ports oropenings, recesses, and the like in the cured material. According tosome embodiments, voids 1818 are filled such that sacrificial layer isflush or even with a surface of initially cured liquid material 1826. Insome embodiments, excess sacrificial layer is removed such that thesurface of initially cured liquid material 1826 is exposed.

Next, a second liquid material 1824 is introduced onto the surface ofinitially cured liquid material 1826 and sacrificial layer 1820. Asdescribed above, a mask 1810 having pattern 1812, a second pattern 1814,a precision light source, or the like is then delivered to the secondliquid material 1824 such that a predetermined portion of second liquidmaterial 1824 becomes photo-activated and cured. Space between thephoto-mask and the previous layer dictates the height of the secondlayer. During the photo-activating and curing process, second liquidmaterial 1824 can also chemically bind to initially cured liquidmaterial 1804, thereby adhering the two layers together and forming asingle device.

The above described process can be repeated to form a multiple layeredpredetermined three-dimensional structure. Following fabrication of thedesired structure, any sacrificial layer 1820 is removed from thestructure, such as by heating or treating the structure with a solvent.The sacrificial layer 1820 removal process, can in some embodiments, beanalogous to negative photoresist chemistry; unexposed regions areremoved and exposed regions remain after solvent rinsing. In someembodiments, the subsequent layers can have the same or differentphysical configuration and/or the same or different material compositionand/or properties, according to desired uses of the device.

In some embodiments, a shape or recess 1806 can be included in chamber1800. Accordingly, when first liquid material 1804 is cured around shape1806, shape will leave a void 1816 in cured first liquid material 1826after the cured first liquid material 1826 is removed from chamber 1800.In some embodiments, shape 1806 can be a chamber, path, system ofchambers, valve ports, or the like commonly associated with microfluidicdevices.

According to yet other embodiments, the base materials of the device caninclude, as disclosed herein, thermally curable components. According tosuch embodiments, following fabrication of the device by build-up ofmultiple layers, as described above, the device can be treated with athermal cure. Thermal cure activates the thermally curable components ofthe base material and further adheres the multiple layers together orcan adhere the device to a second device.

According to some embodiments, the material 1804 of the device caninclude moieties such as moieties disclosed herein, for example asdescribed above with respect to the fourth precursor. According to suchembodiments the moiety precursor remains available at the surfaces ofthe cured device such that a sample passed over the surface can reactwith the moieties. Many functions performed in microfluidic devices relyupon interactions between fluids and transported chemical or biologicalcompounds, therefore, fabricating microfluidic devices with functionalmoiety precursors that remain available after fabrication of the devicecan facilitate microfluidic reactions. According to some embodiments,the moieties included in the material 1804 can be selected and alteredfor a desired function of the fabricated device. Often, spatiallycontrolled surface properties that differ from the bulk materialproperties are desirable in microfluidic devices. Incorporation of aliving radical initiator, in some embodiments for example, in themonomer formulations can enable spatially resolved grafting ormodification via photolithographic methods and surface-mediated livingradical photo-polymerization.

All references cited herein are incorporated herein by reference intheir entirety, including all references cited therein.

EXAMPLES

The following Examples have been included to provide guidance to one ofordinary skill in the art for practicing representative embodiments ofthe presently disclosed subject matter. In light of the presentdisclosure and the general level of skill in the art, those of skill canappreciate that the following Examples are intended to be exemplary onlyand that numerous changes, modifications, and alterations can beemployed without departing from the scope of the presently disclosedsubject matter.

General Considerations

A PFPE microfluidic device has been previously reported by Rolland, J.et al. JACS 2004, 126, 2322-2323, which is incorporated herein byreference in its entirety. The specific PFPE material disclosed inRolland, J., et al., was not chain extended and therefore did notpossess the multiple hydrogen bonds that are present when PFPEs arechain extended with a diisocyanate linker. Nor did the material possesthe higher molecular weights between crosslinks that are needed toimprove mechanical properties such as modulus and tear strength whichare critical to a variety of microfluidics applications. Furthermore,this material was not functionalized to incorporate various moieties,such as a charged species, a biopolymer, or a catalyst.

Herein is described a variety of methods to address these issues.Included in these improvements are methods which describe chainextension, improved adhesion to multiple PFPE layers and to othersubstrates such as glass, silicon, quartz, and other polymers as well asthe ability to incorporate functional monomers capable of changingwetting properties or of attaching catalysts, biomolecules or otherspecies. Also described are improved methods of curing PFPE elastomerswhich involve thermal free radical cures, two-component curingchemistries, and photocuring using photoacid generators.

Example 1

A liquid PFPE precursor having the structure shown below (where n=2) isblended with 1 wt % of a free radical photoinitiator and poured over amicrofluidics master containing 100-μm features in the shape ofchannels. A PDMS mold is used to contain the liquid in the desired areato a thickness of about 3 mm. The wafer is then placed in a UV chamberand exposed to UV light (λ=365) for 10 minutes under a nitrogen purge.Separately, a second master containing 100-μm features in the shape ofchannels is spin coated with a small drop of the liquid PFPE precursorover top of it at 3700 rpm for 1 minute to a thickness of about 20 μm.The wafer is then placed in a UV chamber and exposed to UV light (λ=365)for 10 minutes under a nitrogen purge. Thirdly, a smooth, flat PFPElayer is generated by drawing a doctor's blade across a small drop ofthe liquid PFPE precursor across a glass slide. The Slide is then placedin a UV chamber and exposed to UV light (λ=365) for 10 minutes under anitrogen purge. The thicker layer is then removed, trimmed, and inletholes are punched through it using a luer stub. The layer is then placedon top of the 20-μm thick layer and aligned in the desired area to forma seal. The layers are then placed in an oven and allowed to heat at120° C. for 2 hours. The thin layer is then trimmed and the adheredlayers are lifted from the master. Fluid inlet holes and outlet holesare punched using a luer stub. The bonded layers are then placed on thefully cured PFPE smooth layer on the glass slide and allowed to heat at120° C. for 15 hours. Small needles can then be placed in the inlets tointroduce fluids and to actuate membrane valves as reported by Unger, M.et al. Science. 2000, 288, 113-6.

Example 2 Thermal Free Radical Glass

A liquid PFPE precursor encapped with methacrylate groups is blendedwith 1 wt % of 2,2-Azobisisobutyronitrile and poured over amicrofluidics master containing 100-μm features in the shape ofchannels. A PDMS mold is used to contain the liquid in the desired areato a thickness of about 3 mm. The wafer is then placed in an oven at 65°C. for 20 hours under nitrogen purge. The cured layer is then removed,trimmed, and inlet holes are punched through it using a luer stub. Thelayer is then placed on top of a clean glass slide and fluids can beintroduced through the inlet holes.

Example 3 Thermal Free Radical—Partial Cure Layer to Layer Adhesion

A liquid PFPE precursor encapped with methacrylate groups is blendedwith 1 wt % of 2,2-Azobisisobutyronitrile and poured over amicrofluidics master containing 100-μm features in the shape ofchannels. A PDMS mold is used to contain the liquid in the desired areato a thickness of about 3 mm. The wafer is then placed in an oven at 65°C. for 2-3 hours under nitrogen purge. Separately, a second mastercontaining 100-μm features in the shape of channels is spin coated witha small drop of the liquid PFPE precursor over top of it at 3700 rpm for1 minute to a thickness of about 20 μm. The wafer is then placed in anoven at 65° C. for 2-3 hours under nitrogen purge. Thirdly, a smooth,flat PFPE layer is generated by drawing a doctor's blade across a smalldrop of the liquid PFPE precursor across a glass slide. The wafer isthen placed in an oven at 65° C. for 2-3 hours under nitrogen purge. Thethicker layer is then removed, trimmed, and inlet holes are punchedthrough it using a luer stub. The layer is then placed on top of the20-μm thick layer and aligned in the desired area to form a seal. Thelayers are then placed in an oven and allowed to heat at 65° C. for 10hours. The thin layer is then trimmed and the adhered layers are liftedfrom the master. Fluid inlet holes and outlet holes are punched using aluer stub. The bonded layers are then placed on the partially cured PFPEsmooth layer on the glass slide and allowed to heat at 65° C. for 10hours. Small needles can then be placed in the inlets to introducefluids and to actuate membrane valves as reported by Unger, M. et al.Science. 2000, 288, 113-6.

Example 4 Thermal Free Radical—Partial Cure Adhesion to Polyurethane

A photocurable liquid polyurethane precursor containing methacrylategroups is blended with 1 wt % of 2,2-Azobisisobutyronitrile and pouredover a microfluidics master containing 100-μm features in the shape ofchannels. A PDMS mold is used to contain the liquid in the desired areato a thickness of approximately 3 mm. The wafer is then placed in anoven at 65° C. for 2-3 hours under nitrogen purge. Separately, a secondmaster containing 100-μm features in the shape of channels is spincoated with a small drop of the liquid PFPE precursor over top of it at3700 rpm for 1 minute to a thickness of approximately 20 μm. The waferis then placed in an oven at 65° C. for 2-3 hours under nitrogen purge.Thirdly, a smooth, flat PFPE layer is generated by drawing a doctor'sblade across a small drop of the liquid PFPE precursor across a glassslide. The wafer is then placed in an oven at 65° C. for 2-3 hours undernitrogen purge. The thicker layer is then removed, trimmed, and inletholes are punched through it using a luer stub. The layer is then placedon top of the 20-μm thick layer and aligned in the desired area to forma seal. The layers are then placed in an oven and allowed to heat at 65°C. for 10 hours. The thin layer is then trimmed and the adhered layersare lifted from the master. Fluid inlet holes and outlet holes arepunched using a luer stub. The bonded layers are then placed on thepartially cured PFPE smooth layer on the glass slide and allowed to heatat 65° C. for 10 hours. Small needles can then be placed in the inletsto introduce fluids and to actuate membrane valves as reported by Unger,M. et al. Science. 2000, 288, 113-6.

Example 5 Thermal Free Radical—Partial Cure Adhesion toSilicone-containing Polyurethane

A photocurable liquid polyurethane precursor containing PDMS blocks andmethacrylate groups is blended with 1 wt % of 2,2-Azobisisobutyronitrileand poured over a microfluidics master containing 100-μm features in theshape of channels. A PDMS mold is used to contain the liquid in thedesired area to a thickness of approximately 3 mm. The wafer is thenplaced in an oven at 65° C. for 2-3 hours under nitrogen purge.Separately, a second master containing 100-μm features in the shape ofchannels is spin coated with a small drop of the liquid PFPE precursorover top of it at 3700 rpm for 1 minute to a thickness of approximately20 μm. The wafer is then placed in an oven at 65° C. for 2-3 hours undernitrogen purge. Thirdly, a smooth, flat PFPE layer is generated bydrawing a doctor's blade across a small drop of the liquid PFPEprecursor across a glass slide. The wafer is then placed in an oven at65° C. for 2-3 hours under nitrogen purge. The thicker layer is thenremoved, trimmed, and inlet holes are punched through it using a luerstub. The layer is then placed on top of the 20-μm thick layer andaligned in the desired area to form a seal. The layers are then placedin an oven and allowed to heat at 65° C. for 10 hours. The thin layer isthen trimmed and the adhered layers are lifted from the master. Fluidinlet holes and outlet holes are punched using a luer stub. The bondedlayers are then placed on the partially cured PFPE smooth layer on theglass slide and allowed to heat at 65° C. for 10 hours. Small needlescan then be placed in the inlets to introduce fluids and to actuatemembrane valves as reported by Unger, M. et al. Science. 2000, 288,113-6.

Example 6 Thermal Free Radical—Partial Cure Adhesion to PFPE-PDMS BlockCopolymer

A liquid precursor containing both PFPE and PDMS blocks encapped withmethacrylate groups is blended with 1 wt % of 2,2-Azobisisobutyronitrileand poured over a microfluidics master containing 100-μm features in theshape of channels. A PDMS mold is used to contain the liquid in thedesired area to a thickness of approximately 3 mm. The wafer is thenplaced in an oven at 65° C. for 2-3 hours under nitrogen purge.Separately, a second master containing 100-μm features in the shape ofchannels is spin coated with a small drop of the liquid PFPE precursorover top of it at 3700 rpm for 1 minute to a thickness of approximately20 μm. The wafer is then placed in an oven at 65° C. for 2-3 hours undernitrogen purge. Thirdly, a smooth, flat PFPE layer is generated bydrawing a doctor's blade across a small drop of the liquid PFPEprecursor across a glass slide. The wafer is then placed in an oven at65° C. for 2-3 hours under nitrogen purge. The thicker layer is thenremoved, trimmed, and inlet holes are punched through it using a luerstub. The layer is then placed on top of the 20-μm thick layer andaligned in the desired area to form a seal. The layers are then placedin an oven and allowed to heat at 65° C. for 10 hours. The thin layer isthen trimmed and the adhered layers are lifted from the master. Fluidinlet holes and outlet holes are punched using a luer stub. The bondedlayers are then placed on the partially cured PFPE smooth layer on theglass slide and allowed to heat at 65° C. for 10 hours. Small needlescan then be placed in the inlets to introduce fluids and to actuatemembrane valves as reported by Unger, M. et al. Science. 2000, 288,113-6.

Example 7 Thermal Free Radical—Partial Cure Glass Adhesion

A liquid PFPE precursor encapped with methacrylate groups is blendedwith 1 wt % of 2,2-Azobisisobutyronitrile and poured over amicrofluidics master containing 100-μm features in the shape ofchannels. A PDMS mold is used to contain the liquid in the desired areato a thickness of about 3 mm. The wafer is then placed in an oven at 65°C. for 2-3 hours under nitrogen purge. The partially cured layer isremoved from the wafer and inlet holes are punched using a luer stub.The layer is then placed on top of a glass slide treated with a silanecoupling agent, trimethoxysilyl propyl methacrylate. The layer is thenplaced in an oven and allowed to heat at 65° C. for 20 hours,permanently bonding the PFPE layer to the glass slide. Small needles canthen be placed in the inlets to introduce fluids.

Example 8 Thermal Free Radical—Partial Cure PDMS Adhesion

A liquid poly(dimethylsiloxane) precursor poured over a microfluidicsmaster containing 100-μm features in the shape of channels. The wafer isthen placed in an oven at 80° C. for 3 hours. Separately, a secondmaster containing 100-μm features in the shape of channels is spincoated with a small drop of liquid PFPE precursor encapped withmethacrylate units at 3700 rpm for 1 minute to a thickness of about 20μm. The wafer is then placed in an oven at 65° C. for 2-3 hours undernitrogen purge. The PDMS layer is then removed, trimmed, and inlet holesare punched through it using a luer stub. The layer is then treated withan oxygen plasma for 20 minutes followed by treatment with a silanecoupling agent, trimethoxysilyl propyl methacrylate. The treated PDMSlayer is then placed on top of the partially cured PFPE thin layer andheated at 65° C. for 10 hours. The thin layer is then trimmed and theadhered layers are lifted from the master. Fluid inlet holes and outletholes are punched using a luer stub. The bonded layers are then placedon the partially cured PFPE smooth layer on the glass slide and allowedto heat at 65° C. for 10 hours. Small needles can then be placed in theinlets to introduce fluids and to actuate membrane valves as reported byUnger, M. et al. Science. 2000, 288, 113-6.

Example 9 Thermal Free Radical PDMS Adhesion Using SYLGARD 184® andFunctional PDMS

A liquid poly(dimethylsiloxane) precursor is designed such that it canbe part of the base or curing component of SYLGARD 184®. The precursorcontains latent functionalities such as epoxy, methacrylate, or aminesand is mixed with the standard curing agents and poured over amicrofluidics master containing 100-μm features in the shape ofchannels. The wafer is then placed in an oven at 80° C. for 3 hours.Separately, a second master containing 100-μm features in the shape ofchannels is spin coated with a small drop of liquid PFPE precursorencapped with methacrylate units at 3700 rpm for 1 minute to a thicknessof approximately 20 μm. The wafer is then placed in an oven at 65° C.for 2-3 hours under nitrogen purge. The PDMS layer is then removed,trimmed, and inlet holes are punched through it using a luer stub. ThePDMS layer is then placed on top of the partially cured PFPE thin layerand heated at 65° C. for 10 hours. The thin layer is then trimmed andthe adhered layers are lifted from the master. Fluid inlet holes andoutlet holes are punched using a luer stub. The bonded layers are thenplaced on the partially cured PFPE smooth layer on the glass slide andallowed to heat at 65° C. for 10 hours. Small needles can then be placedin the inlets to introduce fluids and to actuate membrane valves asreported by Unger, M. et al. Science. 2000, 288, 113-6.

Example 10 Epoxy/Amine

A two-component liquid PFPE precursor system such as shown belowcontaining a PFPE diepoxy and a PFPE diamine are blended together in astoichiometric ratio and poured over a microfluidics master containing100-μm features in the shape of channels. A PDMS mold is used to containthe liquid in the desired area to a thickness of about 3 mm. The waferis then placed in an oven at 65° C. for 5 hours. The cured layer is thenremoved, trimmed, and inlet holes are punched through it using a luerstub. The layer is then placed on top of a clean glass slide and fluidscan be introduced through the inlet holes.

Example 11 Epoxy/Amine—Excess Adhesion to Glass

A two-component liquid PFPE precursor system such as shown belowcontaining a PFPE diepoxy and a PFPE diamine are blended together in a4:1 epoxy:amine ratio such that there is an excess of epoxy and pouredover a microfluidics master containing 100-μm features in the shape ofchannels. A PDMS mold is used to contain the liquid in the desired areato a thickness of about 3 mm. The wafer is then placed in an oven at 65°C. for 5 hours. The cured layer is then removed, trimmed, and inletholes are punched through it using a luer stub. The layer is then placedon top of a clean glass slide that has been treated with a silanecoupling agent, aminopropyltriethoxy silane. The slide is then heated at65° C. for 5 hours to permanently bond the device to the glass slide.Fluids can then be introduced through the inlet holes.

Example 12 Epoxy/Amine-Excess Adhesion to PFPE Layers

A two-component liquid PFPE precursor system such as shown belowcontaining a PFPE diepoxy and a PFPE diamine are blended together in a1:4 epoxy:amine ratio such that there is an excess of amine and pouredover a microfluidics master containing 100-μm features in the shape ofchannels. A PDMS mold is used to contain the liquid in the desired areato a thickness of about 3 mm. Separately, a second master containing100-μm features in the shape of channels is coated with a small drop ofliquid PFPE precursors blended in a 4:1 epoxy:amine ratio such thatthere is an excess of epoxy units and spin coated at 3700 rpm for 1minute to a thickness of about 20 μm. The wafer is then placed in anoven at 65° C. for 5 hours. The thick layer is then removed, trimmed,and inlet holes are punched through it using a luer stub. The thicklayer is then placed on top of the cured PFPE thin layer and heated at65° C. for 5 hours. The thin layer is then trimmed and the adheredlayers are lifted from the master. Fluid inlet holes and outlet holesare punched using a luer stub. The bonded layers are then placed on aglass slide treated with a silane coupling agent, aminopropyltriethoxysilane and heated in an oven at 65° C. for 5 hours to adhere the deviceto the glass slide. Small needles can then be placed in the inlets tointroduce fluids and to actuate membrane valves as reported by Unger, M.et al. Science. 2000, 288, 113-6.

Example 13 Epoxy/Amine-Excess Adhesion to PDMS Layers

A liquid poly(dimethylsiloxane) precursor is poured over a microfluidicsmaster containing 100-μm features in the shape of channels. The wafer isthen placed in an oven at 80° C. for 3 hours. Separately, a secondmaster containing 100-μm features in the shape of channels is coatedwith a small drop of liquid PFPE precursors blended in, a 4:1epoxy:amine ratio such that there is an excess of epoxy units and spincoated at 3700 rpm for 1 minute to a thickness of about 20 μm. The waferis then placed in an oven at 65° C. for 5 hours. The PDMS layer is thenremoved, trimmed, and inlet holes are punched through it using a luerstub. The layer is then treated with an oxygen plasma for 20 minutesfollowed by treatment with a silane coupling agent, aminopropyltriethoxysilane. The treated PDMS layer is then placed on top of the PFPE thinlayer and heated at 65° C. for 10 hours to adhere the two layers. Thethin layer is then trimmed and the adhered layers are lifted from themaster. Fluid inlet holes and outlet holes are punched using a luerstub. The bonded layers are then placed on a glass slide treated withaminopropyltriethoxy silane and allowed to heat at 65° C. for 10 hours.Small needles can then be placed in the inlets to introduce fluids andto actuate membrane valves as reported by Unger, M. et al. Science.2000, 288, 113-6.

Example 14 Epoxy/Amine-Excess Adhesion to PFPE Layers Attachment of aBiomolecule

A two-component liquid PFPE precursor system such as shown belowcontaining a PFPE diepoxy and a PFPE diamine are blended together in a1:4 epoxy:amine ratio such that there is an excess of amine and pouredover a microfluidics master containing 100-μm features in the shape ofchannels. A PDMS mold is used to contain the liquid in the desired areato a thickness of about 3 mm. Separately, a second master containing100-μm features in the shape of channels is coated with a small drop ofliquid PFPE precursors blended in a 4:1 epoxy:amine ratio such thatthere is an excess of epoxy units and spin coated at 3700 rpm for 1minute to a thickness of about 20 μm. The wafer is then placed in anoven at 65° C. for 5 hours. The thick layer is then removed, trimmed,and inlet holes are punched through it using a luer stub. The thicklayer is then placed on top of the cured PFPE thin layer and heated at65° C. for 5 hours. The thin layer is then trimmed and the adheredlayers are lifted from the master. Fluid inlet holes and outlet holesare punched using a luer stub. The bonded layers are then placed on aglass slide treated with a silane coupling agent, aminopropyltriethoxysilane and heated in an oven at 65° C. for 5 hours to adhere the deviceto the glass slide. Small needles can then be placed in the inlets tointroduce fluids and to actuate membrane valves as reported by Unger, M.et al. Science. 2000, 288, 113-6. An aqueous solution containing aprotein functionalized with a free amine is then flowed through thechannel which is lined with unreacted epoxy moieties, in such a way thatthe channel is then functionalized with the protein.

Example 15 Epoxy/Amine-Excess Adhesion to PFPE Layers, Attachment of aCharged Species

A two-component liquid PFPE precursor system such as shown belowcontaining a PFPE diepoxy and a PFPE diamine are blended together in a1:4 epoxy:amine ratio such that there is an excess of amine and pouredover a microfluidics master containing 100-μm features in the shape ofchannels. A PDMS mold is used to contain the liquid in the desired areato a thickness of about 3 mm. Separately, a second master containing100-μm features in the shape of channels is coated with a small drop ofliquid PFPE precursors blended in a 4:1 epoxy:amine ratio such thatthere is an excess of epoxy units and spin coated at 3700 rpm for 1minute to a thickness of about 20 μm. The wafer is then placed in anoven at 65° C. for 5 hours. The thick layer is then removed, trimmed,and inlet holes are punched through it using a luer stub. The thicklayer is then placed on top of the cured PFPE thin layer and heated at65° C. for 5 hours. The thin layer is then trimmed and the adheredlayers are lifted from the master. Fluid inlet holes and outlet holesare punched using a luer stub. The bonded layers are then placed on aglass slide treated with a silane coupling agent, aminopropyltriethoxysilane and heated in an oven at 65° C. for 5 hours to adhere the deviceto the glass slide. Small needles can then be placed in the inlets tointroduce fluids and to actuate membrane valves as reported by Unger, M.et al. Science. 2000, 288, 113-6. An aqueous solution containing acharged molecule functionalized with a free amine is then flowed throughthe channel which is lined with unreacted epoxy moieties, in such a waythat the channel is then functionalized with the charged molecule.

Example 16 Epoxy/Amine-Partial Cure Adhesion to Glass

A two-component liquid PFPE precursor system such as shown belowcontaining a PFPE diepoxy and a PFPE diamine are blended together in astoichiometric ratio and poured over a microfluidics master containing100-μm features in the shape of channels. A PDMS mold is used to containthe liquid in the desired area to a thickness of about 3 mm. The waferis then placed in an oven at 65° C. for 0.5 hours such that it ispartially cured. The partially cured layer is then removed, trimmed, andinlet holes are punched through it using a luer stub. The layer is thenplaced on a glass slide treated with a silane coupling agent,aminopropyltriethoxy silane, and allowed to heat at 65° C. for 5 hourssuch that it is adhered to the glass slide. Small needles can then beplaced in the inlets to introduce fluids.

Example 17 Epoxy/Amine-Partial Cure Layer to Layer Adhesion

A two-component liquid PFPE precursor system such as shown belowcontaining a PFPE diepoxy and a PFPE diamine are blended together in astoichiometric ratio and poured over a microfluidics master containing100-μm features in the shape of channels. A PDMS mold is used to containthe liquid in the desired area to a thickness of about 3 mm. The waferis then placed in an oven at 65° C. for 0.5 hours such that it ispartially cured. The partially cured layer is then removed, trimmed, andinlet holes are punched through it using a luer stub. Separately, asecond master containing 100-μm features in the shape of channels isspin coated with a small drop of the liquid PFPE precursors over top ofit at 3700 rpm for 1 minute to a thickness of about 20 μm. The wafer isthen placed in an oven at 65° C. for 0.5 hours such that it is partiallycured. The thick layer is then placed on top of the 20-μm thick layerand aligned in the desired area to form a seal. The layers are thenplaced in an oven and allowed to heat at 65° C. for 1 hour to adhere thetwo layers. The thin layer is then trimmed and the adhered layers arelifted from the master. Fluid inlet holes and outlet holes are punchedusing a luer stub. The bonded layers are then placed on a glass slidetreated with a silane coupling agent, aminopropyltriethoxy silane, andallowed to heat at 65° C. for 10 hours. Small needles can then be placedin the inlets to introduce fluids and to actuate membrane valves asreported by Unger, M. et al. Science. 2000, 288, 113-6.

Example 18 Epoxy/Amine-Partial Cure PDMS Adhesion

A liquid poly(dimethylsiloxane) precursor is poured over a microfluidicsmaster containing 100-μm features in the shape of channels. The wafer isthen placed in an oven at 80° C. for 3 hours. The cured PDMS layer isthen removed, trimmed, and inlet holes are punched through it using aluer stub. The layer is then treated with an oxygen plasma for 20minutes followed by treatment with a silane coupling agent,aminopropyltriethoxy silane. Separately, a second master containing100-μm features in the shape of channels is spin coated with a smalldrop of liquid PFPE precursors mixed in a stoichiometric ratio at 3700rpm for 1 minute to a thickness of about 20 μm. The wafer is then placedin an oven at 65° C. for 0.5 hours. The treated PDMS layer is thenplaced on top of the partially cured PFPE thin layer and heated at 65°C. for 1 hour. The thin layer is then trimmed and the adhered layers arelifted from the master. Fluid inlet holes and outlet holes are punchedusing a luer stub. The bonded layers are then placed on a glass slidetreated with aminopropyltriethoxy silane and allowed to heat at 65° C.for 10 hours. Small needles can then be placed in the inlets tointroduce fluids and to actuate membrane valves as reported by Unger, M.et al. Science. 2000, 288, 113-6.

Example 19 Photocuring with Latent Functional Groups Available Post CureAdhesion To Glass

A liquid PFPE precursor having the structure shown below (where R is anepoxy group, the curvy lines are PFPE chains, and the circle is alinking molecule) is blended with 1 wt % of a free radicalphotoinitiator and poured over a microfluidics master containing 100-μmfeatures in the shape of channels. A PDMS mold is used to contain theliquid in the desired area to a thickness of about 3 mm. The wafer isthen placed in a UV chamber and exposed to UV light (λ=365) for 10minutes under a nitrogen purge. The fully cured layer is then removedfrom the master and inlet holes are punched using a luer stub. Thedevice is placed on a glass slide treated with a silane coupling agent,aminopropyltriethoxy silane, and allowed to heat at 65° C. for 15 hourspermanently bonding the device to the glass slide. Small needles canthen be placed in the inlets to introduce fluids.

Example 20 Photocuring with Latent Functional Groups Available Post CureAdhesion to PFPE

A liquid PFPE precursor having the structure shown below (where R is anepoxy group), the curvy lines are PFPE chains, and the circle is alinking molecule) is blended with 1 wt % of a free radicalphotoinitiator and poured over a microfluidics master containing 100-μmfeatures in the shape of channels. A PDMS mold is used to contain theliquid in the desired area to a thickness of about 3 mm. The wafer isthen placed in a UV chamber and exposed to UV light (λ=365) for 10minutes under a nitrogen purge. The fully cured layer is then removedfrom the master and inlet holes are punched using a luer stub.Separately a second master containing 100-μm features in the shape ofchannels is spin coated with a small drop of the liquid PFPE precursor(where R is an amine group) over top of it at 3700 rpm for 1 minute to athickness of about 20 μm. The wafer is then placed in a UV chamber andexposed to UV light (λ=365) for 10 minutes under a nitrogen purge. Thethicker layer is then placed on top of the 20-μm thick layer and alignedin the desired area to form a seal. The layers are then placed in anoven and allowed to heat at 65° C. for 2 hours. The thin layer is thentrimmed and the adhered layers are lifted from the master. Fluid inletholes and outlet holes are punched using a luer stub. The bonded layersare then placed on a glass slide treated with a silane coupling agent,aminopropyltriethoxy silane, and allowed to heat at 65° C. for 15 hourspermanently bonding the device to the glass slide. Small needles canthen be placed in the inlets to introduce fluids and to actuate membranevalves as reported by Unger, M. et al. Science. 2000, 288, 113-6.

Example 21 Photocuring w/ Latent Functional Groups Available Post CureAdhesion to PDMS

A liquid poly(dimethylsiloxane) precursor is poured over a microfluidicsmaster containing 100-μm features in the shape of channels. The wafer isthen placed in an oven at 80° C. for 3 hours. The cured PDMS layer isthen removed, trimmed, and inlet holes are punched through it using aluer stub. The layer is then treated with an oxygen plasma for 20minutes followed by treatment with a silane coupling agent,aminopropyltriethoxy silane. Separately a second master containing100-μm features in the shape of channels is spin coated with a smalldrop of the liquid PFPE precursor (where R is an epoxy) over top of itat 3700 rpm for 1 minute to a thickness of about 20 μm. The wafer isthen placed in a UV chamber and exposed to UV light (λ=365) for 10minutes under a nitrogen purge. The thicker PDMS layer is then placed ontop of the 20-μm thick layer and aligned in the desired area to form aseal. The layers are then placed in an oven and allowed to heat at 65°C. for 2 hours. The thin layer is then trimmed and the adhered layersare lifted from the master. Fluid inlet holes and outlet holes arepunched using a luer stub. The bonded layers are then placed on a glassslide treated with a silane coupling agent, aminopropyltriethoxy silane,and allowed to heat at 65° C. for 15 hours permanently bonding thedevice to the glass slide. Small needles can then be placed in theinlets to introduce fluids and to actuate membrane valves as reported byUnger, M. et al. Science. 2000, 288, 113-6.

Example 22 Latent Functional Groups Available Post Cure: AttachBiomolecule

A liquid PFPE precursor having the structure shown below (where R is anamine group), the curvy lines are PFPE chains, and the circle is alinking molecule) is blended with 1 wt % of a free radicalphotoinitiator and poured over a microfluidics master containing 100-μmfeatures in the shape of channels. A PDMS mold is used to contain theliquid in the desired area to a thickness of about 3 mm. The wafer isthen placed in a UV chamber and exposed to UV light (λ=365) for 10minutes under a nitrogen purge. The fully cured layer is then removedfrom the master and inlet holes are punched using a luer stub.Separately a second master containing 100-μm features in the shape ofchannels is spin coated with a small drop of the liquid PFPE precursor(where R is an epoxy group) over top of it at 3700 rpm for 1 minute to athickness of about 20 μm. The wafer is then placed in a UV chamber andexposed to UV light (λ=365) for 10 minutes under a nitrogen purge. Thethicker layer is then placed on top of the 20-μm thick layer and alignedin the desired area to form a seal. The layers are then placed in anoven and allowed to heat at 65° C. for 2 hours. The thin layer is thentrimmed and the adhered layers are lifted from the master. Fluid inletholes and outlet holes are punched using a luer stub. The bonded layersare then placed on a glass slide treated with a silane coupling agent,aminopropyltriethoxy silane, and allowed to heat at 65° C. for 15 hourspermanently bonding the device to the glass slide. Small needles canthen be placed in the inlets to introduce fluids and to actuate membranevalves as reported by Unger, M. et al. Science. 2000, 288, 113-6. Anaqueous solution containing a protein functionalized with a free amineis then flowed through the channel which is lined with unreacted epoxymoieties, in such a way that the channel is then functionalized with theprotein.

Example 23 Latent Functional Groups Available Post Cure: Attach ChargedSpecies

A liquid PFPE precursor having the structure shown below (where R is anamine group), the curvy lines are PFPE chains, and the circle is alinking molecule) is blended with 1 wt % of a free radicalphotoinitiator and poured over a microfluidics master containing 100-μmfeatures in the shape of channels. A PDMS mold is used to contain theliquid in the desired area to a thickness of about 3 mm. The wafer isthen placed in a UV chamber and exposed to UV light (λ=365) for 10minutes under a nitrogen purge. The fully cured layer is then removedfrom the master and inlet holes are punched using a luer stub.Separately a second master containing 100-μm features in the shape ofchannels is spin coated with a small drop of the liquid PFPE precursor(where R is an epoxy group) over top of it at 3700 rpm for 1 minute to athickness of about 20 μm. The wafer is then placed in a UV chamber andexposed to UV light (λ=365) for 10 minutes under a nitrogen purge. Thethicker layer is then placed on top of the 20-μm thick layer and alignedin the desired area to form a seal. The layers are then placed in anoven and allowed to heat at 65° C. for 2 hours. The thin layer is thentrimmed and the adhered layers are lifted from the master. Fluid inletholes and outlet holes are punched using a luer stub. The bonded layersare then placed on a glass slide treated with a silane coupling agent,aminopropyltriethoxy silane, and allowed to heat at 65° C. for 15 hourspermanently bonding the device to the glass slide. Small needles canthen be placed in the inlets to introduce fluids and to actuate membranevalves as reported by Unger, M. et al. Science. 2000, 288, 113-6. Anaqueous solution containing a charged molecule functionalized with afree amine is then flowed through the channel which is lined withunreacted epoxy moieties, in such a way that the channel is thenfunctionalized with the charged molecule.

Example 24 Photocuring with Functional Monomers Available Post CureAdhesion to Glass

A liquid PFPE dimethacrylate precursor or a monomethacrylate PFPEmacromonomer is blended with a monomer having the structure shown below(where R is an epoxy group) and blended with 1 wt % of a free radicalphotoinitiator and poured over a microfluidics master containing 100-μmfeatures in the shape of channels. A PDMS mold is used to contain theliquid in the desired area to a thickness of about 3 mm. The wafer isthen placed in a UV chamber and exposed to UV light (λ=365) for 10minutes under a nitrogen purge. The fully cured layer is then removedfrom the master and inlet holes are punched using a luer stub. Thedevice is placed on a glass slide treated with a silane coupling agent,aminopropyltriethoxy silane, and allowed to heat at 65° C. for 15 hourspermanently bonding the device to the glass slide. Small needles canthen be placed in the inlets to introduce fluids.

Example 25 Photocuring with Functional Monomers Available Post CureAdhesion to PFPE

A liquid PFPE dimethacrylate precursor is blended with a monomer havingthe structure shown below (where R is an epoxy group) and blended with 1wt % of a free radical photoinitiator and poured over a microfluidicsmaster containing 100-μm features in the shape of channels. A PDMS moldis used to contain the liquid in the desired area to a thickness ofabout 3 mm. The wafer is then placed in a UV chamber and exposed to UVlight (λ=365) for 10 minutes under a nitrogen purge. The fully curedlayer is then removed from the master and inlet holes are punched usinga luer stub. Separately a second master containing 100-μm features inthe shape of channels is spin coated with a small drop of the liquidPFPE precursor plus functional (where R is an amine group) over top ofit at 3700 rpm for 1 minute to a thickness of about 20 μm. The wafer isthen placed in a UV chamber and exposed to UV light (λ=365) for 10minutes under a nitrogen purge. The thicker layer is then placed on topof the 20-μm thick layer and aligned in the desired area to form a seal.The layers are then placed in an oven and allowed to heat at 65° C. for2 hours. The thin layer is then trimmed and the adhered layers arelifted from the master. Fluid inlet holes and outlet holes are punchedusing a luer stub. The bonded layers are then placed on a glass slidetreated with a silane coupling agent, aminopropyltriethoxy silane, andallowed to heat at 65° C. for 15 hours permanently bonding the device tothe glass slide. Small needles can then be placed in the inlets tointroduce fluids and to actuate membrane valves as reported by Unger, M.et al. Science. 2000, 288, 113-6.

Example 26 Photocuring with Functional Monomers Available Post CureAdhesion to PDMS

A liquid poly(dimethylsiloxane) precursor is poured over a microfluidicsmaster containing 100-μm features in the shape of channels. The wafer isthen placed in an oven at 80° C. for 3 hours. The cured PDMS layer isthen removed, trimmed, and inlet holes are punched through it using aluer stub. The layer is then treated with an oxygen plasma for 20minutes followed by treatment with a silane coupling agent,aminopropyltriethoxy silane. Separately a second master containing100-μm features in the shape of channels is spin coated with a smalldrop of a liquid PFPE dimethacrylate precursor plus functional monomer(where R is an epoxy) plus a photoinitiator over top of it at 3700 rpmfor 1 minute to a thickness of about 20 μm. The wafer is then placed ina UV chamber and exposed to UV light (λ=365) for 10 minutes under anitrogen purge. The thicker PDMS layer is then placed on top of the20-μm thick layer and aligned in the desired area to form a seal. Thelayers are then placed in an oven and allowed to heat at 65° C. for 2hours. The thin layer is then trimmed and the adhered layers are liftedfrom the master. Fluid inlet holes and outlet holes are punched using aluer stub. The bonded layers are then placed on a glass slide treatedwith a silane coupling agent, aminopropyltriethoxy silane, and allowedto heat at 65° C. for 15 hours permanently bonding the device to theglass slide. Small needles can then be placed in the inlets to introducefluids and to actuate membrane valves as reported by Unger M. et al.Science. 2000, 288, 113-6.

Example 27 Photocuring with Functional Monomers Available Post CureAttachment of a Biomolecule

A liquid PFPE dimethacrylate precursor is blended with a monomer havingthe structure shown below (where R is an amine group) and blended with 1wt % of a free radical photoinitiator and poured over a microfluidicsmaster containing 100-μm features in the shape of channels. A PDMS moldis used to contain the liquid in the desired area to a thickness ofabout 3 mm. The wafer is then placed in a UV chamber and exposed to UVlight (λ=365) for 10 minutes under a nitrogen purge. The fully curedlayer is then removed from the master and inlet holes are punched usinga luer stub. Separately a second master containing 100-μm features inthe shape of channels is spin coated with a small drop of the liquidPFPE precursor plus functional (where R is an epoxy group) over top ofit at 3700 rpm for 1 minute to a thickness of about 20 μm. The wafer isthen placed in a UV chamber and exposed to UV light (λ=365) for 10minutes under a nitrogen purge. The thicker layer is then placed on topof the 20-μm thick layer and aligned in the desired area to form a seal.The layers are then placed in an oven and allowed to heat at 65° C. for2 hours. The thin layer is then trimmed and the adhered layers arelifted from the master. Fluid inlet holes and outlet holes are punchedusing a luer stub. The bonded layers are then placed on a glass slidetreated with a silane coupling agent, aminopropyltriethoxy silane, andallowed to heat at 65° C. for 15 hours permanently bonding the device tothe glass slide. Small needles can then be placed in the inlets tointroduce fluids and to actuate membrane valves as reported by Unger, M.et al. Science. 2000, 288, 113-6. An aqueous solution containing aprotein functionalized with a free amine is then flowed through thechannel which is lined with unreacted epoxy moieties, in such a way thatthe channel is then functionalized with the protein.

Example 28 Photocuring with Latent Functional Groups Available Post CureAttachment of Charged Species

A liquid PFPE dimethacrylate precursor is blended with a monomer havingthe structure shown below (where R is an amine group) and blended with 1wt % of a free radical photoinitiator and poured over a microfluidicsmaster containing 100-μm features in the shape of channels. A PDMS moldis used to contain the liquid in the desired area to a thickness ofabout 3 mm. The wafer is then placed in a UV chamber and exposed to UVlight (λ=365) for 10 minutes under a nitrogen purge. The fully curedlayer is then removed from the master and inlet holes are punched usinga luer stub. Separately a second master containing 100-μm features inthe shape of channels is spin coated with a small drop of the liquidPFPE precursor plus functional (where R is an epoxy group) over top ofit at 3700 rpm for 1 minute to a thickness of about 20 μm. The wafer isthen placed in a UV chamber and exposed to UV light (λ=365) for 10minutes under a nitrogen purge. The thicker layer is then placed on topof the 20-μm thick layer and aligned in the desired area to form a seal.The layers are then placed in an oven and allowed to heat at 65° C. for2 hours. The thin layer is then trimmed and the adhered layers arelifted from the master. Fluid inlet holes and outlet holes are punchedusing a luer stub. The bonded layers are then placed on a glass slidetreated with a silane coupling agent, aminopropyltriethoxy silane, andallowed to heat at 65° C. for 15 hours permanently bonding the device tothe glass slide. Small needles can then be placed in the inlets tointroduce fluids and to actuate membrane valves as reported by Unger, M.et al. Science. 2000, 288, 113-6. An aqueous solution containing acharged molecule functionalized with a free amine is then flowed throughthe channel which is lined with unreacted epoxy moieties, in such a waythat the channel is then functionalized with the charged molecule.

Example 29 Fabrication of a PFPE Microfluidic Device Using SacrificialChannels

A smooth, flat PFPE layer is generated by drawing a doctor's bladeacross a small drop of the liquid PFPE dimethacrylate precursor across aglass slide. The Slide is then placed in a UV chamber and exposed to UVlight (λ=365) for 10 minutes under a nitrogen purge. A scaffold composedof poly(lactic acid) in the shape of channels is laid on top of theflat, smooth layer of PFPE. A liquid PFPE dimethacrylate precursor iswith 1 wt % of a free radical photoinitiator and poured over thescaffold. A PDMS mold is used to contain the liquid in the desired areato a thickness of about 3 mm. The apparatus is then placed in a UVchamber and exposed to UV light (λ=365) for 10 minutes under a nitrogenpurge. The device can then be heated at 150° C. for 24 hours to degradethe poly(lactic acid) thus revealing voids left in the shape ofchannels.

Example 30 Adhesion of a PFPE Device to Glass Using 185-nm Light

A liquid PFPE dimethacrylate precursor is blended with 1 wt % of a freeradical photoinitiator and poured over a microfluidics master containing100-μm features in the shape of channels. A PDMS mold is used to containthe liquid in the desired area to a thickness of about 3 mm. The waferis then placed in a UV chamber and exposed to UV light (λ=365) for 10minutes under a nitrogen purge. Separately a second master containing100-μm features in the shape of channels is spin coated with a smalldrop of the liquid PFPE precursor over top of it at 3700 rpm for 1minute to a thickness of about 20 μm. The wafer is then placed in a UVchamber and exposed to UV light (λ=365) for 10 minutes under a nitrogenpurge. The thicker layer is then removed, trimmed, and inlet holes arepunched through it using a luer stub. The layer is then placed on top ofthe 20-μm thick layer and aligned in the desired area to form a seal.The layers are then placed in an oven and allowed to heat at 120° C. for2 hours. The thin layer is then trimmed and the adhered layers arelifted from the master. Fluid inlet holes and outlet holes are punchedusing a luer stub. The bonded layers are then placed on a clean, glassslide in such a way that it forms as seal. The apparatus is exposed to185 nm UV light for 20 minutes, forming a permanent bond between thedevice and the glass. Small needles can then be placed in the inlets tointroduce fluids and to actuate membrane valves as reported by Unger, M.et al. Science. 2000, 288, 113-6.

Example 31 “Epoxy Casing” Method to Encapsulate Devices

A liquid PFPE dimethacrylate precursor is blended with 1 wt % of a freeradical photoinitiator and poured over a microfluidics master containing100-μm features in the shape of channels. A PDMS mold is used to containthe liquid in the desired area to a thickness of about 3 mm. The waferis then placed in a UV chamber and exposed to UV light (λ=365) for 10minutes under a nitrogen purge. Separately a second master containing100-μm features in the shape of channels is spin coated with a smalldrop of the liquid PFPE precursor over top of it at 3700 rpm for 1minute to a thickness of about 20 μm. The wafer is then placed in a UVchamber and exposed to UV light (λ=365) for 10 minutes under a nitrogenpurge. The thicker layer is then removed, trimmed, and inlet holes arepunched through it using a luer stub. The layer is then placed on top ofthe 20-μm thick layer and aligned in the desired area to form a seal.The layers are then placed in an oven and allowed to heat at 120° C. for2 hours. The thin layer is then trimmed and the adhered layers arelifted from the master. Fluid inlet holes and outlet holes are punchedusing a luer stub. The bonded layers are then placed on a clean, glassslide in such a way that it forms as seal. Small needles can then beplaced in the inlets to introduce fluids and to actuate membrane valvesas reported by Unger, M. et al. Science. 2000, 288, 113-6. The entireapparatus can then be encased in a liquid epoxy precursor which ispoured over the device allowed to cure. The casing serves tomechanically bind the device the substrate.

Example 32 Fabrication of a PFPE Device from a Three-Armed PFPEPrecursor

A liquid PFPE precursor having the structure shown below (where thecircle represents a linking molecule) is blended with 1 wt % of a freeradical photoinitiator and poured over a microfluidics master containing100-μm features in the shape of channels. A PDMS mold is used to containthe liquid in the desired area to a thickness of about 3 mm. The waferis then placed in a UV chamber and exposed to UV light (λ=365) for 10minutes under a nitrogen purge. Separately a second master containing100-μm features in the shape of channels is spin coated with a smalldrop of the liquid PFPE precursor over top of it at 3700 rpm for 1minute to a thickness of about 20 μm. The wafer is then placed in a UVchamber and exposed to UV light (λ=365) for 10 minutes under a nitrogenpurge. Thirdly a smooth, flat PFPE layer is generated by drawing adoctor's blade across a small drop of the liquid PFPE precursor across aglass slide. The Slide is then placed in a UV chamber and exposed to UVlight (λ=365) for 10 minutes under a nitrogen purge. The thicker layeris then removed, trimmed, and inlet holes are punched through it using aluer stub. The layer is then placed on top of the 20-μm thick layer andaligned in the desired area to form a seal. The layers are then placedin an oven and allowed to heat at 120° C. for 2 hours. The thin layer isthen trimmed and the adhered layers are lifted from the master. Fluidinlet holes and outlet holes are punched using a luer stub. The bondedlayers are then placed on the fully cured PFPE smooth layer on the glassslide and allowed to heat at 120° C. for 15 hours. Small needles canthen be placed in the inlets to introduce fluids and to actuate membranevalves as reported by Unger, M. et al. Science. 2000, 288, 113-6.

Example 33 Photocured PFPE/PDMS Hybrid

A master containing 100-μm features in the shape of channels is spincoated with a small drop of the liquid PFPE dimethacrylate precursorcontaining photoinitiator over top of it at 3700 rpm for 1 minute to athickness of about 20 μm. A PDMS dimethacrylate containingphotoinitiator is then poured over top of the thin PFPE layer to athickness of 3 mm. The wafer is then placed in a UV chamber and exposedto UV light (λ=365) for 10 minutes under a nitrogen purge. The layer isthen removed, trimmed, and inlet holes are punched through it using aluer stub. The hybrid device is then placed on a glass slide and a sealis formed. Small needles can then be placed in the inlets to introducefluids.

Example 34 Microfluidic Device Formed From Blended Thermally andPhotoCurable Materials

Firstly, a predetermined amount, e.g., 5 grams, of a chain-extended PFPEdimethacrylate containing a small amount of photoinitiator, such ashydroxycyclohexylphenyl ketone, is measured. Next, a 1:1 ratio byweight, e.g., 5 grams, of a chain-extended PFPE diisocyanate is added.Next, an amount, e.g., 0.3 mL, of a PFPE tetrol (Mn˜2000 g/mol) is thenadded such that there is a stoichiometric amount of —N(C═O)— and —OHmoieties. The three components are then mixed thoroughly and degassedunder vacuum.

Master templates are generated using photolithography and are coatedwith a thin layer of metal, e.g., Gold/Palladium, using an argon plasma.Thin layers for devices are spin coated at 1500 rpm from the PFPE blendonto patterned substrates. A thin, flat (non patterned), layer also isspin coated. Separately, thicker layers are cast onto the metal-coatedmaster templates, typically by pooling the material inside, for example,a PDMS gasket. All layers are then placed in a UV chamber, purged withnitrogen for 10 minutes, and photocured for ten minutes into solidrubbery pieces under a thorough nitrogen purge. The layers can then betrimmed and inlet/outlet holes punched. Next the layers are stacked andaligned in registered positions such that they form a conformal seal.The stacked layers are then heated, at 105° C. for 10 minutes. Theheating step initiates the thermal cure of the thermally curablematerial which is physically entangled in the photocured matrix. Becausethe layers are in conformal contact, strong adhesion is obtained. Thetwo adhered layers can then be peeled from the patterned master orlifted off with a solvent, such as dimethyl formamide, and placed incontact with a third flat, photocured substrate which has not yet beenexposed to heat. The three-layer device is then baked for 15 hours at110° C. to fully adhere all three layers.

According to another embodiment, the thermal cure is activated at atemperature of between about 20 degrees Celsius (C) and about 200degrees C. According to yet another embodiment, the thermal cure isactivated at a temperature of between about 50 degrees Celsius (C) andabout 150 degrees C. Further still, the thermal cure selected such thatit is activated at a temperature of between about 75 degrees Celsius (C)and about 200 degrees C.

According to yet another embodiment, the amount of photocure substanceadded to the material is substantially equal to the amount of thermalcure substance. In a further embodiment, the amount of thermal curesubstance added to the material is about 10 percent of the amount ofphotocure substance. According to another embodiment, the amount ofthermal cure substance is about 50 percent of the amount of thephotocure substance.

Example 35 Multicomponent Material for Fabricating Microfluidic Devices

The chemical structure of each component will be described below. In thefollowing example, the first component (Component 1) is a chainextended, photocurable PFPE liquid precursor. The synthesis consists ofthe chain extension of a commercially available PFPE diol (ZDOL) with acommon diisocyanate, isophorone diisocyanate (IPDI), using classicurethane chemistry with an organo-tin catalyst. Following chainextension, the chain is end-capped with a methacrylate-containingdiisocyanate monomer (EIM).

The second component is a chain-extended PFPE diisocyanate. It is madeby the reaction of ZDOL with IPDI in a molar ratio such that theresulting polymer chain is capped with isocyanate groups (Component 2a).The reaction again makes use of classic urethane chemistry with anorgano-tin catalyst.

The second part of the thermally curable component is a commerciallyavailable perfluoropolyether tetraol with a molecular weight of 2,000g/mol (Component 2b).

It will be understood that various details of the presently disclosedsubject matter can be changed without departing from the scope of thepresently disclosed subject matter. Furthermore, the foregoingdescription is for the purpose of illustration only, and not for thepurpose of limitation.

Example 36 Microfluidic Device Formed From Blended Thermally andPhotoCurable Materials: UV Diurethane Methacrylate w/ PFPE Triol+PFPEDiisocyanate System

Firstly, a predetermined amount, for example, 10 grams, of achain-extended PFPE dimethacrylate containing a small amount ofphotoinitiator, such as diethoxyphenyl ketone, is measured. Next, about7 grams, of a PFPE diisocyanate is added. Next, an amount, e.g., 3.0 gof a PFPE triol (Mn˜5000 g/mol) is then added such that there is astoichiometric amount of —N(C═O)— and —OH moieties. The three componentsare then mixed thoroughly and degassed under vacuum.

Master templates are generated using photolithography and are coatedwith a thin layer of metal, e.g., Gold/Palladium, using an Argon plasma.Alternatively the masters are treated with a silane or other releaselayer. Thin layers for devices are spin coated at 1500 rpm from the PFPEblend onto patterned substrates. A thin, flat (non patterned), layeralso is spin coated. Separately, thicker layers are cast onto themetal-coated master templates, typically by pooling the material inside,for example, a PDMS gasket. All layers are then placed in a UV chamber,purged with nitrogen for 10 minutes, and photocured for ten minutes intosolid rubbery pieces under a thorough nitrogen purge. The layers canthen be trimmed and inlet/outlet holes punched. Next the layers arestacked and aligned in registered positions such that they form aconformal seal. The stacked layers are then heated, at 115° C. for 10minutes. The heating step initiates the thermal cure of the thermallycurable material which is physically entangled in the photocured matrix.Because the layers are in conformal contact, strong adhesion isobtained. The two adhered layers can then be peeled from the patternedmaster or lifted off with a solvent, such as dimethyl formamide, andplaced in contact with a third flat, photocured substrate which has notyet been exposed to heat. The three-layer device is then baked for 2hours at 150° C. to fully adhere all three layers.

Example 37 Microfluidic Device Formed From Blended Thermally andPhotoCurable Materials: UV Diurethane Methacrylate w/ PFPE Diepoxy+PFPEDiamine

Firstly, a predetermined amount, e.g., 20 grams, of a chain-extendedPFPE dimethacrylate containing a small amount of photoinitiator, such asdiethoxyphenyl ketone, is measured. Next, about 10 grams, of a PFPEdiepoxy is added. Next, an amount, e.g., 3.6 g of a PFPE diamine is thenadded such that there is a stoichiometric amount of epoxy and aminemoieties. The three components are then mixed thoroughly and degassedunder vacuum.

Master templates are generated using photolithography and are coatedwith a thin layer of metal, e.g., Gold/Palladium, using an Argon plasma.Alternatively, the masters are treated with a silane or other releaselayer. A thin, flat (non patterned), layer is spin coated at 1500 rpm.Separately, thicker layers are cast onto the patterned master templates,typically by pooling the material inside, for example, a PDMS gasket.All layers are then placed in a UV chamber, purged with nitrogen for 10minutes, and photocured for ten minutes into solid rubbery pieces undera thorough nitrogen purge. The layers can then be trimmed andinlet/outlet holes punched. Next the layers are stacked and aligned inregistered positions such that they form a conformal seal. The stackedlayers are then heated, at 120° C. for 20 minutes. The heating stepinitiates the thermal cure of the thermally curable material which isphysically entangled in the photocured matrix. Because the layers are inconformal contact, strong adhesion is obtained.

Example 38 Microfluidic Device Formed From Two Blended PhotoCurableMaterials: UV Diurethane Methacrylate w/ UV Epoxy

Firstly, a predetermined amount, e.g., 10 grams, of a chain-extendedPFPE dimethacrylate containing a small amount of photoinitiator, such asdiethoxyphenyl ketone, is measured. Next, 10 grams, of a PFPE diepoxyformulation which contains 5 grams of ZDOL TX and 0.2 g a photoacidgenerator, such as Rhodorsil 2079, is added. The two components are thenmixed thoroughly and degassed under vacuum.

Master templates are generated using photolithography and are coatedwith a thin layer of metal, e.g., Gold/Palladium, using an Argon plasma.Alternatively, the masters are treated with a silane or other releaselayer. A thin, flat (non patterned), layer is spin coated at 1500 rpm.Separately, thicker layers are cast onto the patterned master templates,typically by pooling the material inside, for example, a PDMS gasket.All layers are then placed in a UV chamber, purged with nitrogen for 5minutes, and photocured for 5 minutes into solid rubbery pieces under athorough nitrogen purge. The layers can then be trimmed and inlet/outletholes punched. Next the layers are stacked and aligned in registeredpositions such that they form a conformal seal. The stacked layers arethen exposed to UV light for a second time for 10 minutes. The second UVstep further propagates the cure of the PFPE diepoxy material which isphysically entangled in the PFPE diurethane methacrylate. Because thelayers are in conformal contact, strong adhesion is obtained.

Example 39 Microfluidic Device Formed From Blended Thermally andPhotoCurable Materials: UV Diurethane Methacrylate w/ PFPE DiisocyanateSystem

Firstly, about 10 grams, of a chain-extended PFPE dimethacrylatecontaining a small amount of photoinitiator, such as diethoxyphenylketone, can be measured. Next, about 7 grams of a PFPE diisocyanateshall be added. The three components shall then be mixed thoroughly anddegassed under vacuum.

Master templates will then be generated using photolithography andcoated with a thin layer of metal, e.g., Gold/Palladium, using an Argonplasma. Alternatively the masters can be treated with a silane or otherrelease layer. Thin layers for devices will then be spin coated at 1500rpm from the PFPE blend onto patterned substrates. A thin, flat (nonpatterned), layer also will be spin coated. Separately, thicker layerscan be cast onto the metal-coated master templates, typically by poolingthe material inside, for example, a PDMS gasket. All layers will then beplaced in a UV chamber, purged with nitrogen for 10 minutes, andphotocured for ten minutes into solid rubbery pieces under a thoroughnitrogen purge. The layers can then be trimmed and inlet/outlet holespunched, as needed. Next the layers will be stacked and aligned inregistered positions such that a conformal seal is formed. The stackedlayers will then be heated, at 130° C. for 30 minutes. The heating stepinitiates thermal cure components of the thermally curable materialwhich are physically entangled in the photocured matrix. Because thelayers are in conformal contact, strong adhesion will be obtained. Thetwo adhered layers can then be peeled from the patterned master orlifted off with a solvent, such as dimethyl formamide, and placed incontact with a third flat, photocured substrate which has not yet beenexposed to heat. The three-layer device can then be baked for 4 hours at130° C. to fully adhere all three layers.

Example 40 Multicomponent Material for Fabricating Microfluidic DevicesMicrofluidic Device Formed From Blended Thermally and PhotoCurableMaterials: UV Diurethane Methacrylate w/ PFPE Diisocyanate System

Firstly, about 10 grams, of a chain-extended PFPE dimethacrylatecontaining a small amount of photoinitiator, such as diethoxyphenylketone, shall be measured. Next, 7 grams of a PFPE diisocyanate shall beadded. The three components will then be mixed thoroughly and degassedunder vacuum.

Master templates will be generated using photolithography and coatedwith a thin layer of metal, e.g., Gold/Palladium, using an argon plasma.Alternatively the masters can be treated with a silane or other releaselayer. Thin layers for devices will then be spin coated at 1500 rpm fromthe PFPE blend onto patterned substrates. A thin, flat (non patterned),layer will also be spin coated. Separately, thicker layers are cast ontothe metal-coated master templates, typically by pooling the materialinside, for example, a PDMS gasket. All layers shall then be placed in aUV chamber, purged with nitrogen for 10 minutes, and photocured for tenminutes into solid rubbery pieces under a thorough nitrogen purge. Thelayers can then be trimmed and inlet/outlet holes punched, as necessary.Next the layers shall be stacked and aligned in registered positionssuch that they form a conformal seal. The stacked layers will then beheated, at 90° C. in a humidity chamber with greater than 50% relativehumidity for 30 minutes (moisture cure). The heating step initiates thethermal cure of the thermally curable material which is physicallyentangled in the photocured matrix. Because the layers are in conformalcontact, strong adhesion will be obtained. The two adhered layers canthen be peeled from the patterned master or lifted off with a solvent,such as dimethyl formamide, and placed in contact with a third flat,photocured substrate which has not yet been exposed to heat. Thethree-layer device can then be baked at 90° C. in a humidity chamberwith greater than 50% relative humidity for 4 hours.

1. A polymer composition, comprising: a first component including afluoropolymer having a first curable functional group, wherein thefluoropolymer having a first curable functional group comprises adiurethane methacrylate functionalized perfluoropolyether having one ofthe following structures:

wherein n′ is an integer from 1 to 100, and n is one; and a secondcomponent including a polymer having a second curable functional group,wherein the polymer having said second curable functional group is afluoropolymer, wherein said fluoropolymer comprises: (a) a diisocyanatefunctionalized perfluoropolyether having a structure of:

wherein n′ is an integer from 1 to 100; (b) a diisocyanatefunctionalized perfluoropolyether having a structure of:

wherein n′ is an integer from 1 to 100; (c) a diepoxy functionalizedperfluoropolyether having a structure of:

wherein n′ is an integer from 1 to 100; or (d) a diamine functionalizedperfluoropolyether having a structure of:

wherein n′ is an integer from 1 to
 100. 2. The polymer composition ofclaim 1, further comprising a third component including a fluoropolymerhaving a third curable functional group, wherein said third curablefunctional group comprises a tetrol functionalized perfluoropolyetherhaving a structure of:

wherein n′ is an integer from 1 to 100; or a triol functionalizedperfluoropolyether having a structure of:

wherein n′ is an integer from 1 to
 100. 3. The composition of claim 2,wherein the first curable functional group reacts at a first wavelengthand the second curable functional group reacts at a second time period,wherein following activation of the second curable functional group thecomposition can be bound to a substrate through activation of the thirdcurable functional group.
 4. The composition of claim 2, wherein thesecond curable functional group reacts at a first time period and thethird curable functional group reacts at a second time period, whereinfollowing activation of the second curable functional group thecomposition can be bound to a substrate through activation of the thirdcurable functional group.
 5. The composition of claim 2, wherein thesecond curable functional group reacts at a first temperature and thethird curable functional group reacts at a second temperature, whereinfollowing activation of the second curable functional group thecomposition can be bound to a substrate through activation of the thirdcurable functional group.
 6. The polymer composition of claim 1, whereinthe polymeric material has a surface energy of less than about 20dynes/cm.
 7. The polymer composition of claim 1, wherein the polymericmaterial has a surface energy of less than about 18 dynes/cm.
 8. Thepolymer composition of claim 1, wherein the polymeric material has asurface energy of less than about 15 dynes/cm.
 9. The composition ofclaim 1, wherein the second curable functional group remains viableafter the first curable functional group is activated such that thesecond curable functional group can bind with a group consisting ofanother polymer, a hydroxyl group, another functional group, andcombinations thereof.
 10. The composition of claim 1, wherein theperfluoropolyether has a molecular weight of about 16000 and a modulusof about 800 kPa.
 11. The composition of claim 1, wherein theperfluoropolyether has a molecular weight of less than about
 16000. 12.The composition of claim 1, wherein the perfluoropolyether has a modulusof greater than about 500 kPa.
 13. The composition of claim 1, whereinthe perfluoropolyether has a molecular weight of about 16000 and apercent elongation at break of about 200 percent.
 14. The composition ofclaim 1, wherein the perfluoropolyether has a percent elongation atbreak of less than about 200 percent.
 15. The composition of claim 1,wherein one of the first component or the second component includes afluoropolymer having an elongation at break of about 300 percent. 16.The composition of claim 1, wherein one of the first component or thesecond component includes a fluoropolymer having an elongation at breakof about 200 percent.
 17. The composition of claim 1, wherein one of thefirst component or the second component includes a fluoropolymer havingan elongation at break of between about 100 percent and about 300percent.
 18. The composition of claim 1, wherein one of the firstcomponent or the second component includes a perfluoropolyether havingan elongation at break of between about 200 percent and about 300percent.